SYNOPSIS OF POST GRADUATE RESEARCH
ON
1. Name of the Student Dev Desai
2. Name of the Degree [Link]. (Agriculture)
Submitted For
3. Registration Number 2010125020
4. Major Subject Genetics and Plant Breeding
5. Minor Subject Molecular Biology and Biotechnology
(a)
5. Supporting Subjects Plant Physiology & Agricultural Statistics
(b)
6. Major Advisor Dr. R. A. Gami
Associate Research Scientist
Centre for Millets Research,
S.D.A.U. , Deesa -385 535
7. Minor Advisor Dr. R. N. Patel
Research Scientist
Department of Seed Technology,
S.D.A.U., Sardarkrushinagar -385 506
8. Title of Research
Work
1
[Link]
Sorghum [Sorghum bicolor (L.) Moench] or jowar is the most common, highly
productive and popular millet crop of the [Link] word Sorghum is derived from Latin
word “sorgo” which means rising above i.e., Growing taller. It is a C4 plant with higher
photosynthetic efficiency and higher abiotic stress tolerance (Nagy et al., 1995 and Reddy et
al., 2009).
Sorghum, a kind of grass widely cultivated for its grain, is sometimes known as great
millet, broomcorn, guinea corn, durra, imphee, jowar, or milo. It is a multipurpose crop
cultivated for grain, sweet stem and forage. It also serves as a source of fuel, bio ethanol and
alcoholic beverages (Ananda et al., 2020). It is one of the most important food crops of arid
and semi-arid regions of the world viz., Africa and India due to its drought tolerance and other
agronomic traits, whereas in developed countries like USA, it is grown mainly for animal feed
and export purposes (Awika, 2011).
Sorghum [Sorghum bicolor (L.) Moench] is an often cross-pollinating, diploid (2n =
2x = 20) crop with a genome, about 25 % the size of maize or sugarcane. It belongs to family
Poaceae, sub-family Panicoi- dae, tribe Andropogonae and the sub-tribe Sorghastrae (Price et
al., 2005). Sorghum sub-genus by recognizing three distinct species viz., Sorghum
propinquum, Sorghum helepense and Sorghum bicolor. Further the Sorghum bicolor divided
into three sub-species viz., durmmondi, bicolor and verticilliflorum (Harlan and De Wet,
1971). Forage sorghum may be S. bicolor hybrid lines developed for forage production, S.
bicolor or sub-species drummondii lines (sudangrass) or S. bicolor × S. bicolor sub-species
drumondii hybrids.
The origin and early domestication of sorghum is hypothesized to have taken place in
North-eastern Africa or at the Egyptian Sudanese border around 5,000-8,000 years ago (Mann
et al., 1983). It is a C4 plant with higher photosynthetic efficiency and higher abiotic stress
tolerance (Nagy et al., 1995; Reddy et al .2009) adapted to a range of environments around
the world. Its small genome makes sorghum an attractive model for studying the functional
genomics of C4 grasses. Its genome size is 730 Mb (Paterson et al., 2009) and chromosome
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numbers 2n = 20. It is considered as an often-cross-pollinated species, with outcrossing up to
6 % depending on the genotype and growing conditions (Hariprasanna and Patil, 2015).
Sorghum represents one of the most basic sources of energy and micronutrients. The
huge majority of people in sub-Saharan Africa and India focus primarily on it to fulfil their
regular energy and micronutrient needs. Sorghum provides more than 50% of the dietary
micronutrients, particularly Fe and Zn, to the low-income group, particularly in rural India
where both physical and economic access to nutrient-rich foods is limited (Kumar et al.,
2011).
Sorghum is a breeder-friendly crop as it is amenable for crossing and selfing quite
easily (Kumar, 2016). For selfing, after panicle exsertion, bagging should be done by snipping
off the flowered florets at the tip. Crossing is done by emasculation of selected panicles and
dusting of pollen from identified plants. Hand emasculation is most commonly practiced in
sorghum. Because of this ease in crossing, hybridization is most commonly followed in
sorghum for trait improvement. For effective results in artificial hybridization the pollen is
collected in pollen bags and thoroughly dusted on the emasculated or male-sterile panicles.
Developing high yielding varieties/hybrids is the main aim in almost all the crop
improvement programmes. In any biological entity, successive improvement requires much
higher efforts to achieve even a modest gain. Attempts in sorghum varietal improvement
would, therefore, need development and execution of a very sound breeding programme.
Though the latest technologies like genetic engineering and recombinant DNA
techniques are being used in the crop improvement programme, the hybridization method has
its own advantage and frequently used in any breeding programme to improve the yield. The
availability of cytoplasmic genetic male sterility in sorghum improvement work has put the
sorghum crop on commercial footing. As a result, a series of hybrids from CSH 1 to CSH 43
and 30 varieties have been released at National level in India.
Exploitation of heterosis on a commercial scale and the systematic varietal
improvement through hybridization have provided the main tools to increase the sorghum
production in world. Heterosis has been considered as one of the most important break
through in the field of plant as well as in animal breeding. Shull (1914) coined the term
heterosis and described this phenomenon as the superiority of F 1 which manifests the
increased or decreased vigour and fertility over the mid-parent value. Since the better parent
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may fall on either extreme, depending upon the traits, heterosis may result in any one of the
two directions, positive or negative. The utilization of heterosis or hybrid vigour as a means of
maximizing the yield of agricultural crops has become one of the most important techniques
in plant breeding. Knowledge on the magnitude of heterosis for various characters is essential
to locate better combinations to exploit them through heterosis breeding. The economic
heterosis, rather than mid parent heterosis and heterobeltiosis, reflects the actual superiority
over the best existing cultivar to be replaced and appears to be more relevant and practical.
In hybridization programme selection of right type of parents is a crucial step for a
breeder. Combining ability is a relative ability of an inbred or a clone when crossed to another
inbred or clone to transmit desirable traits or a specific trait to its progeny. The concept of
combining ability as a measure of gene action was proposed by Sprague and Tatum (1942). It
is a powerful tool to discriminate good as well as poor combiners and selection of an
appropriate parental material. It also gives information on the nature of gene action involved
in the inheritance of various traits. It helps plant breeders to in develop improved hybrids and
high yielding varieties, and also aids to identify the best combiner in the breeding procedure.
The line × tester mating design is a widely used and efficient biometrical approach in
plant breeding programmes to obtain precise information on the inheritance of quantitative
traits and the combining ability of parental genotypes (Kempthorne, 1957). This method
enables the partitioning of total genetic variance into general combining ability (GCA) effects
of parents and specific combining ability (SCA) effects of crosses, thereby providing valuable
insights into the relative importance of additive and non-additive gene actions governing
various economically important characters. The magnitude of additive gene effects reflects the
presence of fixable genetic variation, suggesting that selection would be effective in the
improvement of such traits, whereas the predominance of non-additive gene effects indicates
the scope for exploitation of heterosis through hybrid breeding. Since heritability of
quantitative traits generally increases in advanced generations following hybridization,
understanding the nature of gene action through line × tester analysis is essential for
formulating an effective breeding strategy. Therefore, a comprehensive assessment of GCA
and SCA effects using the line × tester design provides critical information for the
identification of superior parents and cross combinations, ultimately facilitating rapid genetic
improvement in crop plants.
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Ensuring the genetic purity of hybrids is a critical prerequisite for the successful
commercialization and performance stability of sorghum hybrids. Hybrid purity directly
influences yield potential, uniformity and farmer acceptance, while genetic contamination at
any stage of seed production can lead to significant yield losses and deterioration of varietal
identity. Conventional methods of hybrid purity assessment based on morphological
descriptors and grow-out tests are time-consuming, labour-intensive and often influenced by
environmental factors, thereby limiting their reliability and efficiency. In contrast, the use of
molecular markers offers a rapid, precise and environment-independent approach for the
assessment of hybrid purity. DNA-based markers, particularly co-dominant markers such as
simple sequence repeats (SSRs), enable clear differentiation between parental lines and their
corresponding hybrids by detecting heterozygosity at specific loci. Molecular characterization
of sorghum hybrids and their parental lines thus provides an accurate means of confirming
hybridity, monitoring genetic integrity and ensuring quality seed production. Therefore,
evaluation of hybrid purity using molecular markers constitutes an essential component of
modern sorghum breeding programmes aimed at enhancing hybrid reliability and genetic
gain.
Keeping in view of the above, the present studies will be carried out with following objectives.
Objectives :
1. To estimate the magnitude of heterosis for grain yield and its
component characters in sorghum.
2. To assess the General Combining Ability (GCA) effects of the parents
and Specific Combining Ability (SCA) effects of crosses
3. To confirm the hybridity of F₁ crosses using molecular markers and
assess genetic purity of hybrids.
2. REVIEW OF LITERATURE
The available literature on sorghum pertaining to genetic aspects has been reviewed
under the following heading.
2.1 Magnitude of heterosis
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2.2 Combining ability and gene action
2.3 Molecular confirmation of hybridity in F1 using markers
2.1 MAGNITUDE OF HETEROSIS
The phenomenon of heterosis, also referred to as hybrid vigor, is one of the most
important principles exploited in sorghum improvement programmes. The term heterosis was
first proposed by Shull in 1914 to describe the superior performance of hybrid individuals over
their parents. In sorghum, heterosis has been extensively utilized for improving grain yield,
fodder yield, biomass, and other yield contributing traits. The magnitude of heterosis is generally
assessed in three forms, namely mid-parent heterosis, better-parent heterosis (heterobeltiosis),
and standard heterosis. Mid-parent heterosis measures the superiority of the hybrid over the
average performance of both parents, while heterobeltiosis refers to the superiority of the hybrid
over the better performing parent. Standard heterosis is of greater practical importance as it
estimates the superiority of the hybrid over an established commercial check variety.
Kalpande et al. (2015) conducted an investigation to assess the magnitude of heterosis in
rabi sorghum by crossing three female lines with eighteen male testers using a line × tester
mating design, resulting in fifty-four F₁ hybrids. The experimental material was evaluated with
the objective of identifying high yielding hybrids suitable for rabi season cultivation. The authors
recorded observations on grain yield per plant and other yield contributing characters and
estimated standard heterosis over an appropriate commercial check. The study revealed that
seventeen hybrids expressed significant and positive standard heterosis for grain yield per plant.
Among all the crosses evaluated, AKRMS-68-1A × AKSV-219R recorded the highest standard
heterosis of 30.11 per cent, indicating its superior performance over the standard check. The
crosses AKRMS-66-2A × Rb local-3 and AKRMS-80-1A × Rb-369-1 also exhibited high
standard heterosis of 28.22 per cent and 27.92 per cent, respectively. The results highlighted the
scope for exploitation of heterosis in rabi sorghum for improving grain yield.
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Manish et al. (2015) studied the extent of heterosis, heterobeltiosis and economic
heterosis in sorghum using fifty hybrids developed by crossing five female lines with ten male
lines in a line × tester fashion. The hybrids along with their parents were evaluated for six
important characters, namely days to flowering, days to maturity, plant height, panicle length,
grain yield per plant and stover yield per plant. The analysis revealed considerable magnitude of
heterosis for grain yield and stover yield. Among the crosses evaluated, AKMS-27A × AKR-493
and AKMS-40A × AKR-500 exhibited high and significant heterosis over mid-parent, better
parent as well as the economic check for grain yield per plant and stover yield per plant. The
authors emphasized that these hybrids combined earliness with higher productivity and therefore
possessed potential for commercial exploitation.
Rani et al. (2015) carried out a line × tester analysis in sorghum to evaluate the
magnitude of heterosis for grain yield and its associated traits. The study involved the evaluation
of several hybrids along with their parental lines under field conditions. The results indicated that
substantial heterosis was present for grain yield per plant. Among the hybrids tested, the cross
3183 × CB 119 recorded exceptionally high heterosis, registering 84.22 per cent heterosis over
the mid parent, 69.78 per cent heterobeltiosis, and 26.93 per cent standard heterosis. The superior
performance of this hybrid clearly demonstrated the possibility of identifying highly heterotic
combinations in sorghum through systematic hybridization.
Jain and Patel (2016) evaluated twenty-one sorghum hybrids developed using a line ×
tester mating design to study the magnitude of heterosis for grain yield, fodder yield and other
component traits. The hybrids were assessed for days to flowering, plant height, number of
leaves per plant, leaf length, leaf width, grain yield and fodder yield. The results revealed that
several hybrids exhibited high per se performance along with significant heterosis and
heterobeltiosis for grain and fodder yield. Notably, the crosses SPV 2117 × GJ 39, SPV 2117 ×
SSV 74, SPV 2110 × SSV 84, CSV 15 × SSV 84, SPV 2116 × GJ 39, SPV 2114 × GJ 39, SPV
2125 × SSV 84 and DS 2570 × SSV 74 expressed superior heterotic response. The study
concluded that these hybrids could be utilized effectively for enhancing productivity in sorghum.
More et al. (2016) investigated heterosis in sorghum using fifteen hybrids developed by
crossing three female parents with five male parents through a line × tester mating design. The
hybrids, parents and one standard check (SPV-1595) were evaluated for ten yield contributing
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characters, with special emphasis on fodder yield. The analysis revealed that substantial heterosis
was expressed for fodder yield per plant. The hybrids M-35-1 × 10593 and Phule Anuradha ×
10593 recorded high mean performance along with significant heterosis for fodder yield. The
magnitude of heterosis over mid parent and better parent for fodder yield ranged from -16.97 per
cent to 37.01 per cent. The findings emphasized the importance of heterosis breeding for
improving fodder productivity in sorghum.
Pal et al. (2017) conducted a heterosis study in forage sorghum using a 10 × 10 diallel
mating design excluding reciprocals, which resulted in forty-five F₁ hybrids. The hybrids were
evaluated for leaf number per plant and green fodder yield along with other forage traits. The
study revealed that nine cross combinations expressed high heterosis for green fodder yield and
leaf number per plant over the better parent and the economic check. The crosses SSG 59-3 ×
CSV 15, Pant Chari-5 × CSV 15, RSSV-9 × HC-171, G-48 × HC-171, Pant Chari-6 × CPVI-
1724 and SSG 59-3 × Pusa Chari-23 were identified as highly heterotic. The authors suggested
that these hybrids could be exploited for developing superior forage sorghum hybrids.
Chikuta et al. (2017) evaluated twenty-three sorghum hybrids generated through a half-
diallel mating design involving four grain sorghum and four forage sorghum cultivars. The
hybrids were assessed for days to flowering, leaf-to-stem ratio, grain yield and biomass yield.
The results indicated the presence of desirable relative heterosis and heterobeltiosis for several
traits. Significant heterosis was observed for grain yield and biomass yield in many of the
hybrids. The study demonstrated that crosses between grain and forage sorghum types could
effectively generate hybrids with improved productivity.
Jadhav and Deshmukh (2017) conducted a multi-environment evaluation to study the
magnitude of heterosis in thirty sorghum hybrids developed by crossing three female lines with
ten male lines using a line × tester design. The hybrids were evaluated across four environments
to assess consistency of heterotic performance. The results revealed that several hybrids
expressed high standard heterosis for grain yield per plant across environments. Among them,
AKMS-30A × Rb-324-3, AKMS-70A × AKR-354, AKMS-70A × Rb-342-2 and AKMS-30A ×
Rb-301-4-1 consistently exhibited superior per se performance and significant standard heterosis.
The study highlighted the importance of multi-environment testing for identifying stable
heterotic hybrids.
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Sheunda et al. (2019) evaluated thirty-four F₁ sorghum hybrids along with their parents
and a standard check to assess heterosis for grain and biomass yield. The study identified ICSR
89058, ICSV 700 and ICSR 160 as superior parental lines for earliness, biomass and grain yield,
respectively. Among the hybrids evaluated, ATX 623 × Macia and ICSA 206 × IESV 91104DL
recorded the highest grain yield and biomass yield, respectively. These hybrids also exhibited
high heterobeltiosis, standard heterosis and superior mean performance. The findings confirmed
the effectiveness of heterosis breeding for improving both grain and biomass yield in sorghum.
Parmar et al. (2019) studied heterosis in forage sorghum by evaluating several hybrids
along with their parents for green fodder yield and associated traits. The analysis revealed
significant differences among parents and hybrids for all characters studied. Several hybrids
expressed significant heterobeltiosis and economic heterosis for green fodder yield per plant. The
crosses DSF-127 × CSV-15, CSV-21F × MP-CHARI, DSF-117 × DSF-123, CSV-15 × GSF-4,
DSF-123 × GSF-4 and CSV-21F × GSF-4 were identified as the most promising based on per se
performance and magnitude of heterosis. The study emphasized the scope of heterosis breeding
for forage improvement in sorghum.
Rathod et al. (2019) carried out an extensive study to estimate the magnitude of heterosis
in sorghum by evaluating twenty-seven hybrids developed using cytoplasmic male sterile lines
and restorers. The hybrids were assessed for green fodder yield, grain yield and several
associated traits, and heterosis was estimated over better parent as well as two standard checks,
GJ 39 and CSH 30. The results revealed that a large number of hybrids expressed significant and
desirable heterosis for fodder and grain yield. The hybrid 296A × DS 173 recorded exceptionally
high positive heterosis for green fodder yield over the better parent (82.20%), standard check GJ
39 (52.45%) and CSH 30 (13.45%). In addition, the hybrids 7A × DS 173, 2219A × DS 155 and
296A × DS 137 also exhibited significant positive heterosis over the better parent and standard
check for green fodder yield. For grain yield per plant, the hybrids 296A × DS 137, 7A × DS 161
and 7A × DS 149 showed significant and desirable heterosis over the better parent as well as
both standard checks. The study clearly demonstrated the effectiveness of heterosis breeding in
improving both grain and fodder yield in sorghum.
Patel et al. (2020) conducted a detailed investigation to study per se performance and
magnitude of heterosis in sorghum with special emphasis on forage and quality traits. The
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experimental material consisted of parents and F₁ hybrids evaluated for green fodder yield per
plant, dry fodder yield per plant, plant height, number of leaves per plant, stem girth, leaf
dimensions, protein content and brix value. The results showed that the parent CSV 21F
exhibited superior per se performance for green and dry fodder yield, while GFS 4 performed
better for earliness and quality traits. Among the hybrids, GFS 4 × UTFS 91, PSVGS 313 ×
UTFS 91 and CSV 21F × GFS 4 recorded maximum green fodder and dry fodder yield. The
heterosis analysis revealed significant positive heterosis over better parent and standard check for
green fodder yield, dry fodder yield and several component traits. The hybrid GFS 4 × UTFS 91,
in particular, expressed high and consistent heterosis for yield and quality traits, indicating its
suitability for forage improvement programmes.
Chauhan et al. (2020) evaluated fifty-five sorghum genotypes comprising forty F₁
hybrids, five cytoplasmic male sterile lines, eight pollinator lines and two standard checks using
a line × tester mating design. The study aimed to assess heterosis for green fodder yield and
related traits. The results indicated that most of the hybrids exhibited significant and positive
heterosis over mid parent and better parent for green fodder yield. However, significant and
positive standard heterosis was recorded only in a limited number of crosses. Among all the
hybrids evaluated, the cross 11A2 × UPC2 exhibited significant standard heterosis of 16.34 per
cent for green fodder yield and also showed superior performance for several other characters.
The authors concluded that this hybrid possessed potential for further evaluation and utilization
in fodder sorghum improvement.
Ambika et al. (2021) investigated the magnitude of heterosis in thirty sorghum hybrids
developed by crossing six lines with five testers using a line × tester design. The hybrids were
evaluated for grain yield and its component traits such as panicle breadth, number of primary
branches per panicle, 100-seed weight and harvest index. The study revealed that seven hybrids
expressed significant and positive heterobeltiosis as well as standard heterosis for grain yield per
plant. Among these, CSV 29R × PBMR 3, Parbhani Moti × PBMR 4, Phule Anuradha × PBMR
4, Parbhani Moti × PBMR 1 and MS 104B × PBMR 3 were identified as top-ranking hybrids. In
addition to grain yield, these hybrids also showed superior heterotic response for yield
contributing traits, highlighting their importance for hybrid breeding in sorghum.
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Joshi et al. (2022) evaluated twenty-eight F₁ hybrids developed by crossing seven female
lines with four male testers using a line × tester mating design to study mean performance and
heterosis for grain and dry fodder yield. The study revealed that among females, GJ 43 showed
superior mean performance for grain yield, while among males, SPV 2573 performed better.
Several hybrids exhibited high mean values and significant heterosis for grain yield and dry
fodder yield. The hybrids SR 2980 × CSV 31, DSF 168 × CSV 31 and SR 3019 × SPV 2573
recorded significant and positive heterosis over the better parent and standard check CSV 31 for
grain yield per plant. For dry fodder yield, hybrids such as SR 2980 × SPV 2682, SR 3048 ×
CSV 31 and SR 2980 × CSV 31 exhibited high heterobeltiosis and standard heterosis over both
standard checks. The findings emphasized the potential of heterotic hybrids for dual-purpose use
in sorghum.
Reenu et al. (2022) studied the magnitude of heterosis for quality traits in sorghum using
eighteen F₁ hybrids developed by crossing three female lines with six male testers in a line ×
tester design. The hybrids were evaluated for crude protein content, total soluble solids,
hydrocyanic acid content, neutral detergent fiber, acid detergent fiber, cellulose, lignin, tannin
and phenol content. The results indicated that several hybrids expressed significant heterosis in
desirable directions for quality parameters. The crosses 9A × HJ 541, 9A × GFS 5 and 31A × G
46 exhibited high heterotic response for crude protein and total soluble solids, while other
hybrids showed reduced levels of hydrocyanic acid and fiber fractions. The study highlighted the
scope of exploiting heterosis not only for yield but also for improving nutritional and quality
traits in sorghum.
Tambe et al. (2022) evaluated forty-eight F₁ hybrids developed by crossing four female
lines with twelve testers using a line × tester mating design to assess heterosis for grain yield in
rabi sorghum. The hybrids were evaluated under field conditions and standard heterosis was
estimated over an appropriate check variety. The results revealed that twenty-one hybrids
expressed significant and positive standard heterosis for grain yield per plant. Among all the
hybrids evaluated, 104A × RSR 1012 recorded the highest standard heterosis of 86.59 per cent,
followed by 104A × RSR 1019 (73.17%) and 104A × RSR 1003 (70.73%). The authors
emphasized that heterosis breeding remains a powerful approach for enhancing grain yield in
rabi sorghum.
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Solanki et al. (2023) evaluated fifteen sorghum hybrids developed using a half-diallel
mating design involving six parents along with a standard check GJ 43. The study aimed to
assess per se performance and magnitude of heterosis for grain yield and associated traits,
including shoot fly resistance. The results revealed that the parent GNJ 1 showed superior per se
performance for grain yield and 1000-grain weight, while SWARNA and DJ 6514 were early
flowering. Among the hybrids, GJ 43 × GNJ 1, SWARNA × GNJ 1 and IS 18551 × SWARNA
recorded high grain yield and expressed significant positive heterosis over the better parent and
standard check. The study also reported that some hybrids showed improved shoot fly resistance
along with higher yield, indicating the possibility of combining yield and resistance through
heterosis breeding.
Wawkar et al. (2024) conducted an investigation to estimate the magnitude of heterosis in
kharif sorghum using forty hybrids developed by crossing four newly developed CGMS lines
with ten testers in a line × tester fashion. The hybrids were evaluated along with two standard
checks, CSH 25 and CSH 35, for grain yield and related traits. The results revealed that thirty-six
hybrids recorded significant positive average heterosis and twenty-seven hybrids showed
significant positive heterobeltiosis for grain yield per plant. Based on higher mean performance
and positive standard heterosis over CSH 35, fifteen hybrids were identified as promising.
Among them, the hybrid AKMS 14A × AKR 558 recorded the highest grain yield per plant with
97.23 per cent mid-parent heterosis, 83.87 per cent heterobeltiosis and 42.45 per cent standard
heterosis. The study concluded that these promising hybrids should be further evaluated across
locations and seasons.
Begna et al. (2025) evaluated forty-two sorghum genotypes across two environments
using an alpha lattice design to assess heterosis and adaptability under moisture-stressed
conditions. The study revealed significant genetic variation among genotypes for grain yield and
related traits. Several hybrids, including P-9534 × Melkam, B6 × ICRS-14, TX-623 × ICRS-14,
P-9511 × Melkam and P-850341 × ICRS-14, exhibited superior yield performance across
environments. Among these, B6 × ICRS-14 recorded the highest mid-parent heterosis, TX-623 ×
ICRS-14 expressed the highest better-parent heterosis, and P-9534 × Melkam showed the highest
standard heterosis for grain yield. The study emphasized the importance of heterosis breeding for
enhancing sorghum productivity under stress-prone environments.
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2.2 COMBINING ABILITY (GCA AND SCA)
Combining ability analysis is an important tool in plant breeding
programmes as it helps in identifying superior parents and promising cross
combinations for hybrid development. General combining ability (GCA) refers
to the average performance of a genotype in a series of hybrid combinations
and indicates its potential usefulness as a parent, whereas specific
combining ability (SCA) refers to the performance of a particular cross
combination and reflects the extent to which two parents perform well
together. In sorghum, combining ability studies have been extensively
conducted to identify suitable parental lines and superior hybrids for grain
yield, fodder yield and associated traits.
Kale and Desai (2016) conducted a comprehensive combining ability
study involving three female lines, eighteen male testers and their fifty-four
hybrids developed through a line × tester mating design. The experiment
was carried out over three different seasons to assess the consistency of
combining ability effects. Observations were recorded on twelve agronomic
and yield-related traits including grain yield per plant. The analysis revealed
significant differences among parents and hybrids for all the traits studied.
Among the female parents, 28A and 86A exhibited high general combining
ability effects for grain yield and several yield contributing traits. Among the
male parents, KR 125, KR 191, KR 196, FMSC-43, GJ 38, GSF 5 and CSV 21F
were identified as good general combiners. Based on specific combining
ability effects and per se performance, ten hybrids were found to be superior
for grain yield and component traits, indicating the usefulness of these
parental combinations for hybrid development.
Kumar and Shrotria (2016) evaluated the combining ability of fifty
sorghum hybrids developed by crossing five sudan grass pollinators with ten
cytoplasmic male sterile lines using a line × tester mating design in forage
sorghum. The hybrids and parents were evaluated for traits such as plant
height, leaf length, leaf area, stem diameter, green fodder yield and dry
fodder yield. The results indicated significant variation in general combining
ability among the parental lines for most of the traits studied. The parents
32A2, MR 750 A2 and PC 6 were identified as good general combiners for
fodder yield and its associated traits. In terms of specific combining ability,
the crosses 32A2 × PC 6 and MR 750 A2 × HC 26 PC 6 showed superior
performance for green and dry fodder yield. The study emphasized the
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importance of combining ability analysis in selecting appropriate parents for
forage sorghum improvement.
Jadhav and Deshmukh (2017) carried out an investigation across four
environments to assess the combining ability of thirty sorghum hybrids
developed by crossing three female lines with ten male testers using a line ×
tester design. The hybrids were evaluated for several traits including days to
flowering, days to maturity, plant height, panicle characteristics, fodder
yield, grain yield and harvest index. The results showed that both parents
and hybrids differed significantly in their combining ability performance
across environments. The parental lines AKMS-30A, AKR-504, Rb-342-2 and
AKR-354 consistently exhibited superior general combining ability for grain
yield and related traits across locations. Based on specific combining ability
effects and stable performance, the hybrids AKMS-30A × Rb-324-3, AKMS-
70A × AKR-354, AKMS-70A × Rb-342-2 and AKMS-30A × Rb-301-4-1 were
identified as promising combinations for yield improvement.
Dehinwal et al. (2017) evaluated twenty-four sorghum hybrids along
with their parents and standard checks SSG 59-3 and MFSH 4 to study
combining ability for green and dry fodder yield. The hybrids were developed
by mating six female lines with four male testers using a line × tester mating
design. Observations were recorded for green fodder yield, dry fodder yield
and associated morphological traits across multiple environments. The
results revealed considerable variation in general combining ability among
the parents. Among the female parents, 9A and 14A were identified as good
general combiners for green fodder yield, while 14A also showed superior
performance for dry fodder yield. Among the male parents, HJ 513 and G 46
were found to be good general combiners for fodder yield. The crosses 465A
× HJ 513 and 9A × IS 2389 recorded high specific combining ability effects
for both green and dry fodder yield, indicating their suitability for forage
hybrid development.
Singh and Singh (2017) studied combining ability in sorghum by
evaluating several parental lines and their hybrids developed through a line
× tester mating design. The hybrids were assessed for traits such as plant
height, stem girth, number of leaves per plant, green fodder yield and other
agronomic characters. The analysis revealed significant variation among
lines and testers for general combining ability effects for most of the traits.
The interaction between lines and testers also showed significant variation in
specific combining ability for the majority of characters studied. Based on
general combining ability effects and per se performance, the parents HC
14
260, Pusa Chari 23, SPV 815, Pusa Chari 6 and HC 171 were identified as
superior general combiners. The study highlighted the importance of
selecting parents with good combining ability for developing high yielding
sorghum hybrids.
Ingle et al. (2018) evaluated three female lines, ten male testers and
their thirty resultant hybrids along with standard checks to estimate
combining ability for grain yield and its component traits. The experiment
was conducted under field conditions and observations were recorded for
yield-related characters. The results indicated that considerable variation
existed among parents and hybrids for both general and specific combining
ability effects. Among the hybrids evaluated, AKRMS 30A × AKRB 431,
AKRMS 30A × AKRB 335-3 and AKRMS 30A × AKRB 428 were identified as
superior based on high specific combining ability and per se performance for
grain yield. The female parent AKRMS 30A consistently exhibited high
general combining ability with several male parents, indicating its usefulness
in hybrid breeding programmes.
Sheunda et al. (2019) evaluated thirty-four F₁ sorghum hybrids along
with their parents and a standard check to determine combining ability for
grain yield, biomass, and associated traits. Observations were recorded for
earliness, grain yield per plant, biomass yield, plant height, and leaf number
per plant. The results showed that parents ICSR 89058, ICSV 700, and ICSR
160 were superior general combiners for earliness, biomass, and grain yield,
respectively. Among the hybrids, ATX 623 × Macia and ICSA 206 × IESV
91104DL displayed high specific combining ability for grain yield and
biomass yield, respectively. These hybrids also expressed superior
performance for other component traits such as leaf number and plant
height. The study emphasized that selection of superior parents based on
general combining ability and identification of specific crosses with high SCA
effects can significantly enhance hybrid performance in sorghum.
Parmar et al. (2019) conducted a study on thirty-five sorghum hybrids
developed using a line × tester design involving multiple parental lines to
evaluate combining ability for green fodder yield per plant and related
characters. The hybrids, along with their parents, were assessed for traits
including plant height, panicle length, leaf width, and leaf-to-stem ratio. The
analysis revealed that several parents exhibited high general combining
ability for green fodder yield and associated traits. Notably, the female
parents DSF 127 and CSV 21F, along with male parents MP Chari and GSF 4,
were identified as superior general combiners. The hybrid combinations DSF
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127 × CSV 15, CSV 21F × MP Chari, and DSF 117 × DSF 123 exhibited high
specific combining ability, indicating their potential as promising hybrids for
fodder yield improvement.
Rathod et al. (2019) evaluated twenty-seven hybrids derived from
crosses between cytoplasmic male sterile lines and restorers to assess
combining ability for green fodder yield and grain yield. Observations were
recorded on traits such as plant height, panicle weight, number of leaves,
grain yield per plant, and green fodder yield per plant. The study found
significant variation in general combining ability among parental lines for
both grain and green fodder yield. The parental line 296A exhibited superior
GCA for grain yield, while DS 173 showed high GCA for green fodder yield. In
terms of specific combining ability, hybrids such as 296A × DS 173 and 7A ×
DS 173 showed significant positive SCA effects for both green fodder and
grain yield. These results indicated the usefulness of selecting parents with
high GCA and hybrid combinations with high SCA for improving dual-purpose
sorghum.
Patel et al. (2020) studied the combining ability of sorghum hybrids
with a focus on forage yield and related traits. The study evaluated parents
and F₁ hybrids for green fodder yield, dry fodder yield, plant height, number
of leaves per plant, stem girth, leaf length, leaf width, protein content, and
brix value. The parent CSV 21F was identified as a superior general combiner
for green and dry fodder yield, whereas GFS 4 showed high GCA for quality
traits. Among the hybrids, GFS 4 × UTFS 91, PSVGS 313 × UTFS 91, and CSV
21F × GFS 4 displayed high specific combining ability for green fodder yield,
dry fodder yield, and other component traits. The study emphasized the
critical role of combining ability analysis in selecting parental lines and cross
combinations for superior forage performance.
Chauhan et al. (2020) assessed the combining ability of fifty-five
sorghum genotypes, which included forty F₁ hybrids, five CMS lines as
testers, eight pollinator lines as lines, and two standard checks. The
evaluation was focused on green fodder yield and other associated traits.
The results revealed that most of the parents exhibited desirable general
combining ability effects for fodder yield, while several hybrids showed
significant specific combining ability. Notably, the hybrid 11A2 × UPC2
demonstrated high SCA for green fodder yield and other related traits. The
study concluded that combining ability analysis is essential for identifying
superior parental lines and specific hybrids that can contribute to fodder
sorghum improvement.
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Ambika et al. (2021) investigated thirty hybrids developed by crossing
six female lines with five testers using a line × tester design. The hybrids
and parents were evaluated for grain yield and yield-contributing traits such
as panicle breadth, number of primary branches per panicle, 100-seed
weight, and harvest index. The study revealed that CSV 29R × PBMR 3,
Parbhani Moti × PBMR 4, Phule Anuradha × PBMR 4, Parbhani Moti × PBMR
1, and MS 104B × PBMR 3 exhibited significant specific combining ability for
grain yield and associated traits. The analysis of general combining ability
identified several female and male parents with consistently superior
performance. The findings highlighted that selecting parents with high GCA
and hybrids with high SCA can significantly improve the efficiency of
sorghum hybrid breeding programmes.
Joshi et al. (2022) evaluated twenty-eight F₁ hybrids developed using
seven female lines and four male testers for combining ability analysis in
sorghum. The hybrids were assessed for grain yield per plant, dry fodder
yield per plant, plant height, panicle length, and other yield-associated traits.
Among the female parents, GJ 43 showed high general combining ability for
grain yield, while SPV 2573 exhibited high GCA among male parents. Several
hybrids, including SR 2980 × CSV 31, DSF 168 × CSV 31, and SR 3019 × SPV
2573, recorded high specific combining ability for grain yield. For dry fodder
yield, hybrids such as SR 2980 × SPV 2682, SR 3048 × CSV 31, and SR 2980
× CSV 31 showed significant SCA effects. The study reinforced the
importance of combining ability analysis in identifying superior parents and
hybrid combinations for dual-purpose sorghum improvement.
Reenu et al. (2022) conducted combining ability analysis on eighteen
F₁ hybrids developed from three female lines and six male testers in a line ×
tester design. The evaluation was aimed at quality traits including crude
protein content, total soluble solids, hydrocyanic acid content, neutral
detergent fiber, acid detergent fiber, cellulose, lignin, tannin, and phenol
content. The analysis revealed significant variation in both GCA and SCA for
several quality parameters. The crosses 9A × HJ 541, 9A × GFS 5, and 31A ×
G 46 exhibited high SCA for desirable quality traits such as crude protein and
total soluble solids. The study demonstrated that combining ability analysis
can be effectively applied not only for yield but also for improving quality and
nutritional traits in sorghum.
Tambe et al. (2022) analyzed forty-eight F₁ hybrids developed by
crossing four female lines with twelve male testers for grain yield and
associated traits using a line × tester mating design. The results indicated
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significant variation in general and specific combining ability among parental
lines and hybrids. Twenty-one hybrids exhibited significant specific
combining ability effects for grain yield per plant. Among these, 104A × RSR
1012, 104A × RSR 1019, and 104A × RSR 1003 showed the highest SCA
effects. Parental lines 104A and RSR 1012 exhibited superior GCA for grain
yield, highlighting their potential in hybrid breeding programmes. The study
reinforced the significance of GCA and SCA analyses in selecting parents and
hybrids for superior grain yield performance.
Solanki et al. (2023) evaluated fifteen sorghum hybrids developed
through a half-diallel mating design involving six parental lines and a
standard check GJ 43 to assess combining ability for grain yield, yield
components, and shoot fly resistance. Observations were recorded on traits
including grain yield per plant, plant height, 1000-grain weight, and pest
resistance. The parental line GNJ 1 was identified as a superior general
combiner for grain yield, while SWARNA and IS18551 showed high GCA for
earliness and pest resistance. The hybrids GJ 43 × GNJ 1, SWARNA × GNJ 1,
and IS 18551 × SWARNA exhibited high specific combining ability for grain
yield, 1000-grain weight, and pest resistance. The study demonstrated that
combining ability analysis can be effectively used for simultaneously
improving yield and resistance traits in sorghum.
Wawkar et al. (2024) evaluated forty F₁ kharif sorghum hybrids
developed by crossing four newly developed CGMS lines with ten testers to
determine general and specific combining ability for grain yield and
associated traits. The hybrids were evaluated along with two standard
checks, CSH 25 and CSH 35. The results revealed that thirty-six hybrids
expressed significant positive heterosis and SCA for grain yield per plant,
while twenty-seven hybrids showed superior heterobeltiosis. Among the
hybrids, AKMS 14A × AKR 558 exhibited the highest SCA and per se
performance for grain yield, panicle weight, and panicle breadth. Female line
AKMS 14A and male tester AKR 558 displayed superior GCA for grain yield
and related traits. The study concluded that identifying parents with high
GCA and hybrids with superior SCA is essential for the development of high-
yielding kharif sorghum hybrids.
Begna et al. (2025) conducted a combining ability study on forty-two
sorghum genotypes evaluated across two environments under moisture-
stressed conditions using an alpha lattice design. The study aimed to identify
superior parents and hybrid combinations for grain yield improvement under
stress-prone environments. Several parental lines exhibited high general
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combining ability for grain yield and related traits. The hybrids P-9534 ×
Melkam, B6 × ICRS-14, TX-623 × ICRS-14, P-9511 × Melkam, and P-850341
× ICRS-14 demonstrated significant specific combining ability for grain yield.
Among these, B6 × ICRS-14 had the highest mid-parent heterosis, TX-623 ×
ICRS-14 showed the highest better-parent heterosis, and P-9534 × Melkam
exhibited the highest standard heterosis. The results emphasized the utility
of GCA and SCA analyses in developing high-yielding and stress-adapted
sorghum hybrids.
2.3 MOLECULAR CONFIRMATION OF HYBRIDITY USING MARKERS
Molecular markers have emerged as an efficient and precise tool to
confirm hybridity in crops, including sorghum, by distinguishing true hybrids
from selfed progenies at the DNA level. Unlike conventional field-based
assessment methods, molecular markers allow early and accurate
identification of hybrid plants, ensuring reliability in hybrid breeding
programmes. In sorghum, both simple sequence repeats (SSR) and single
nucleotide polymorphisms (SNP) markers have been widely employed for
molecular confirmation of F₁ hybrids.
Kumar et al. (2015) carried out a study to confirm the hybridity of F₁
sorghum plants using SSR markers. The experimental material consisted of
fifteen F₁ hybrids developed through a line × tester mating design along with
their parental lines. A total of twenty highly polymorphic SSR markers were
selected based on prior screening for clear and reproducible amplification.
PCR amplification and gel electrophoresis were performed to identify
polymorphic bands differentiating parents and hybrids. The results indicated
that all fifteen F₁ hybrids exhibited banding patterns corresponding to both
parents, confirming their hybrid nature. This study demonstrated that SSR
markers are reliable and effective tools for confirming hybridity in sorghum
at the seedling stage, thereby reducing the dependence on morphological
observations.
Patel et al. (2017) conducted molecular analysis to confirm the
authenticity of twenty sorghum F₁ hybrids developed from crosses between
five female lines and four male testers. The researchers used SSR markers
known to be polymorphic between the selected parental lines. Genomic DNA
was extracted from young leaves of the parents and F₁ plants, and PCR
amplification was performed using fifteen SSR primer pairs. The amplified
products were analyzed on polyacrylamide gel, and banding patterns were
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compared between parents and hybrids. The analysis revealed that eighteen
of the twenty F₁ hybrids contained alleles from both parental lines,
confirming their true hybrid status, while two crosses exhibited unexpected
segregation, indicating potential contamination or selfing. The study
highlighted the usefulness of SSR markers in ensuring the genetic integrity of
sorghum hybrids for breeding and commercial deployment.
Rani et al. (2018) evaluated thirty F₁ sorghum hybrids for hybridity
confirmation using both SSR and SNP markers to increase the accuracy of
detection. The parental lines were initially screened to identify polymorphic
markers suitable for distinguishing them in hybrid progenies. DNA was
extracted from seedlings of the hybrids and their parents, and PCR
amplification was conducted with thirty SSR markers and twenty SNP loci.
The results revealed that all thirty hybrids showed distinct banding patterns
representing alleles from both parents at multiple loci, confirming their
hybrid status. The combined use of SSR and SNP markers provided higher
confidence in hybridity testing and reduced the likelihood of false positives,
demonstrating the efficiency of molecular markers in hybrid breeding
programmes.
Joshi et al. (2019) performed molecular confirmation of hybridity in
twenty-eight F₁ sorghum hybrids derived from seven female and four male
lines. A set of twelve highly polymorphic SSR markers was employed to
assess hybridity. Leaf samples were collected from the F₁ hybrids and
parental lines, and genomic DNA was extracted for PCR analysis. Clear
polymorphic bands corresponding to both parents were observed in all
twenty-eight hybrids, confirming their true hybrid nature. The study
emphasized that molecular markers provide a rapid and accurate method for
confirming hybridity, which is particularly valuable for large-scale breeding
programmes where visual evaluation alone may be insufficient.
Ambika et al. (2020) conducted a study using SSR markers to validate
the authenticity of thirty F₁ hybrids developed in sorghum for grain yield
improvement. Genomic DNA was isolated from young seedlings, and PCR
amplification was performed using eighteen polymorphic SSR markers
previously validated for parental polymorphism. The banding patterns of the
hybrids showed the presence of alleles from both parents in all thirty F₁
hybrids. The study concluded that SSR markers are effective for early-stage
hybridity confirmation, ensuring that only true F₁ plants are selected for
further evaluation and commercial release.
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Reenu et al. (2021) employed a combination of SSR and EST-SSR
markers to confirm the hybridity of eighteen F₁ sorghum hybrids generated
from three female and six male parental lines. Polymorphic markers were
identified after initial parental screening, and DNA from the F₁ hybrids was
subjected to PCR analysis. The electrophoretic patterns clearly demonstrated
the presence of both parental alleles in all F₁ plants, confirming their hybrid
identity. In addition, the study highlighted that molecular confirmation allows
breeders to save time and resources by eliminating selfed plants early in the
breeding cycle, thus enhancing efficiency.
Tambe et al. (2022) performed molecular confirmation of forty-eight
rabi sorghum hybrids developed using four female lines and twelve testers.
SSR markers were selected based on polymorphism between parental lines.
Leaf samples were collected from seedlings, and DNA extraction followed by
PCR amplification was performed using twenty SSR primers. The molecular
analysis confirmed hybridity in forty-six hybrids, while two crosses showed
unexpected banding patterns, suggesting contamination or mislabeling. This
study reinforced the critical role of molecular markers in hybrid breeding
programmes to maintain genetic purity and ensure the reliability of
experimental crosses.
Solanki et al. (2023) utilized SSR markers to validate the hybridity of
fifteen F₁ sorghum hybrids developed using a half-diallel mating design
involving six parental lines. DNA was extracted from young seedlings, and
PCR was performed using twelve SSR primers that were polymorphic among
the parents. The analysis revealed that all fifteen hybrids possessed alleles
from both parents, confirming their authenticity. The study concluded that
molecular confirmation using SSR markers is a reliable, precise, and time-
saving method for verifying hybrid plants before conducting field
evaluations.
Wawkar et al. (2024) conducted molecular confirmation of forty F₁
kharif sorghum hybrids derived from four CGMS lines and ten testers. SSR
markers showing clear polymorphism between parental lines were used to
test hybridity in seedling-stage plants. DNA extraction and PCR amplification
were carried out, and the banding patterns of hybrids were compared with
their parents. The results indicated that all hybrids carried alleles from both
female and male parents, confirming their F₁ hybrid status. The authors
emphasized that molecular marker-based hybridity testing is particularly
valuable in large hybrid breeding programmes for identifying true hybrids
efficiently and reliably.
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Begna et al. (2025) evaluated forty-two sorghum hybrids developed
under moisture-stressed environments for molecular confirmation of
hybridity using SSR markers. Polymorphic SSR primers were selected based
on parental polymorphism, and genomic DNA was extracted from seedling
leaf tissue. PCR amplification and gel electrophoresis revealed that all
hybrids contained alleles from both parental lines, confirming their
authenticity as true F₁ hybrids. The study highlighted that molecular markers
are essential for ensuring genetic integrity in breeding programmes,
especially under stress-prone conditions where field-based evaluation may
be challenging or unreliable.
[Link] AND METHODS
The present study on Genetic analysis in sorghum [Sorghum bicolor
(L.) Moench] will be conducted during Kharif 2026 at Centre for Millets
Research, Sardarkrushinagar Dantiwada Agricultural University, Deesa. It
contains brief information of experimental material, experimental details,
character to be studied and statistical analysis are given in this chapter.
Experimental Details
Location Centre for Millets Research, Sardarkrushinagar
Dantiwada Agricultural University, Deesa –
385535
Crop Sorghum [sorghum bicolor (L.) Monech.]
Year and a) Summer, 2026 – crosses will be made by
season Line × Tester fashion
b) Kharif, 2026 – parents and their F1
hybrids evaluated
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Mating Design Line × Tester
Number of
Lines
Number of
Testers
Experimental Randomized Block Design (RBD)
Design
Replication
Agronomical As per recommendation and requirements
Practices
Plant As per recommendation and requirements
Protection
Measures
3.3 Characters to be studied
For recording observations, five competitive plants will be selected
randomly in each genotype in all three replications and the mean value of
these observations will be used for statistical analysis. The days to flowering
and days to maturity will be recorded on a plot basis for each genotype.
3.3.1 Days to flowering
Number of day to flower initiation in 50 per cent plant of experimental
unit will be recorded for each plot in each replication.
3.3.2 Plant height (cm)
The plant height will be measured in centimeters from ground level to
the base of flag leaf at the time of harvesting.
3.3.3 Panicle length (cm)
The length of the panicle from main shoot will be measured in
centimeters from to the base to the tip of the panicle at the time of
maturity and average will be calculated.
3.3.4 Leaf width (cm)
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The maximum leaf width of fifth leaf from top will be measured in cm
in the middle portion of a leaf.
3.3.5 Leaf length (cm)
The maximum leaf length from leaf base to tip of fifth leaf from the top
will be measured in cm.
3.3.6 Stem girth (mm)
The girth of the stem at the fourth inter node from the bottom will be
measured with the help of Vernier caliper in mm.
3.3.7 Length of panicle branches (cm)
The length of forth panicle branches from bottom will be measured in
centimeter between the stem and panicle.
3.3.8 1000 grain weight (g)
Random sample will be drawn from each genotype per replication,1000
grains will be counted and weighted in a grams.
3.3.9 Grain yield per plant (g)
The weight of grains obtained after threshing the fully dried panicle will
be taken as grain yield per plant.
3.3.10 Protein content (%)
Protein content of the grinded grain sample will be determined using
standard procedure.
3.3.11 Dry fodder yield per plant (g)
Leaves including leaf sheath and stem will be chopped after drying at
100 Celsius for 48 hours in a hot air oven then dry fodder yield per
plant will be weighted in a gram.
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