Marker-assisted Backcross Breeding Research Articles

Advances in Breeding for Oil Quality Enhancement in Indian Mustard (Brassica spp. L.): Achievements, Challenges, and Research Opportunities

Major objectives in oil crop improvement are enhancement of seed and oil yield, quality of oil according to its use, i.e. edible or industrial uses, breeding of varieties that fit in different cropping systems and breeding biotic and abiotic stress resistant/tolerant varieties. Despite traditional breeding approaches, including pure line breeding, yielding only modest gains in productivity, recent advancements in mustard breeding have led to significant breakthroughs in both productivity and oil quality. This review discusses the innovative breeding strategies that have contributed to these advancements, with a focus on hybrid development, oil quality enhancement, and biotechnological approaches. To enhance productivity, researchers at the University of Delhi have developed hybrid seed production techniques using transgenic Barnase-barstar systems and cytoplasmic male sterility (CMS) systems. These systems enable large-scale hybrid seed production, with field trials demonstrating significant yield heterosis ranging from 31% to 55% compared to national check varieties. In addition to productivity, improving oil and meal quality has been a key objective. By integrating genes from canola-quality mustard lines, breeders have achieved reductions in erucic acid and glucosinolates, enhancing the health profile and industrial applicability of mustard oil. A high-density linkage map developed using an F1 double haploid mapping population has facilitated the marker-assisted backcross breeding of desirable traits, enabling precise transfer of key quality traits. Transgenic approaches, such as antisense RNA technology, have led to the development of high-oleic, low-linoleic mustard lines with improved fatty acid profiles. These advancements reflect a strategic combination of conventional and biotechnological methods, demonstrating a clear pathway for boosting mustard yields while enhancing oil quality. Molecular markers reported for genetic diversity assessment, mapping and tagging genes/QTLs for different qualitative and quantitative traits and their use in marker-assisted selection have been presented. This progress not only addresses current challenges but also sets the stage for future research aimed at further optimizing productivity, oil quality, and resistance to pests and diseases in mustard cultivation.

Incorporation of Photoperiod Insensitivity and High-Yield Genes into an Indigenous Rice Variety from Myanmar, Paw San Hmwe

Paw San Hmwe (PSH) is an indigenous rice variety from Myanmar with a good taste, a pleasant fragrance, and excellent elongation ability during cooking. However, its low yield potential and strong photoperiod sensitivity reduce its productivity, and it is vulnerable to climate changes during growth. To improve the photoperiod insensitivity, yield, and plant stature of PSH, the high-yield genes Grain number 1a (Gn1a) and Wealthy Farmer’s Panicle (WFP), together with the photoperiod insensitivity trait, were introgressed into PSH via marker-assisted backcross breeding and phenotype selection. For the photoperiod insensitivity trait, phenotypic selection was performed under long-day conditions during the dry season. After foreground selection of Gn1a and WFP via simple sequence repeat genotyping, genotyping-by-sequencing was conducted to validate the introgression of target genes and determine the recurrent parent genome recovery of the selected lines. The improved lines were insensitive to photoperiod, and the Gn1a and WFP introgression lines showed significantly higher numbers of primary panicle branches and spikelets per panicle than the recurrent parent, with comparative similarity in cooking and eating qualities. This study successfully improved PSH by decreasing its photoperiod sensitivity and introducing high-yield genes via marker-assisted selection. The developed lines can be used for crop rotation and double-season cropping of better-quality rice.

Genetic enhancement of reproductive stage drought tolerance in RPHR-1005R and derivative rice hybrids through marker-assisted backcross breeding in rice (Oryza sativa L.).

Drought stress is considered as one of the major production constraints in rice. RPHR-1005R is a restorer line (R-Line) with a popular, medium-slender grain type, and is the male parent of the popular Indian rice hybrid, DRR-H3. However, both the hybrid and its restorer are highly vulnerable to the drought stress, which limits the adoption of the hybrid. Therefore, the selection of the restorer line RPHR-1005R has been made with the objective of enhancing drought tolerance. In this study, we have introgressed a major QTL for grain yield under drought (qDTY 1.1) from Nagina22 through a marker-assisted backcross breeding (MABB) strategy. PCR based SSR markers linked to grain yield under drought (qDTY1.1 - RM431, RM11943), fertility restorer genes (Rf3-DRRM-Rf3-10, Rf4-RM6100) and wide compatibility (S5n allele) were deployed for foreground selection. At BC2F1, a single plant (RPHR6339-4-16-14) with target QTL in heterozygous condition and with the highest recurrent parent genome recovery (85.41%) and phenotypically like RPHR-1005R was identified and selfed to generate BC2F2. Fifty-eight homozygous lines were advanced to BC2F4 and six promising restorer lines and a hybrid combination (APMS6A/RPHR6339-4-16-14-3) were identified. In summary, the six improved restorer lines could be employed for developing heterotic hybrids possessing reproductive stage drought tolerance. The hybrid combination (APMS6A/RPHR6339-4-16-14-3) was estimated to ensure stable yields in drought-prone irrigated lowlands as well as in directly seeded aerobic and upland areas of India.

Development of high-amylose maize (Zea mays L.) genotypes adapted to Indian conditions through molecular breeding

Context Amylose is a type of resistant starch with numerous health benefits and industrial applications. Starch from maize (Zea mays L.) usually has an amylose content of ~25%. Aims The aim was to develop high-amylose maize genotypes suitable for human consumption and adapted to Indian conditions. Methods Marker-assisted backcross breeding was used to transfer the mutant ae1 allele from a high-amylose donor from the USA into the three parents (HKI 1344, HKI 1378, HKI 1348-6-2) of two high-yielding white maize hybrids (HM5 and HM12) grown in India. Key results In converted lines, amylose content was 40.40–58.10% of total kernel starch, compared with 22.25–26.39% in parents. The percentage increase in amylose content was 63.70–153.03%. There was a significant amount of background recovery in each backcross generation: 66.80–79% in BC1F1, 72.85–88.60% in BC2F1, and 84.45–93.70% in BC2F2. Overall, the total kernel starch content was reduced (by ~22%) in the ae1-introgressed families. Conclusions The converted lines developed in the study are enriched with kernel amylose while showing significant background recovery. Implications The high-amylose lines developed may be highly beneficial for diabetic patients and in the bioplastics industry, and should be suitable for growing under Indian conditions.

Genetic Improvement of Drought Tolerance in a Mega-Rice Variety Improved White Ponni through Marker-Assisted Backcross Breeding

In the ever-changing climatic conditions, it has become important to enhance rice productivity to ensure global food security. Drought is one of the major limiting factors in rice pro- duction. Drought during the reproductive stage results in maximum or complete yield loss. Efforts have been taken to develop drought-tolerant rice lines by introgressing three major drought-effect QTLs, viz. qDTY1.1, qDTY2.1, and qDTY3.1, from Apo into a susceptible popular rice variety, Improved White Ponni (IWP). Backcross inbred lines of IWP × Apo were developed through the marker-assisted backcross breeding approach. Foreground analysis using linked markers resulted in the identification of 17 progenies carrying two or more QTLs, and the recurrent parent genome recoveries of these lines were >95.6% using 72 genome-wide SSR markers distributed throughout all chromosomes. Upon phenotypic evaluation of 17 IWP BILs, the water limited condition resulted in the identification of improved lines by recording the yield and the yield-related parameters. The promising performance of IWP BILs in terms of spikelet fertility (63.3%) and grain yield per plant (>10 g) under drought stress indicated the positive effects of introgressed qDTYs, while IWP recorded complete yield loss (94.2%). Out of the cultivars considered, the best-performing lines which truly exhibited drought tolerance, with more increased yield than the recurrent parent under water-limited conditions, and the effects of these QTLs and their interactions were examined in this research work.

Phenotypical and molecular characterization of new pepper genotypes resistant to Chili pepper mild mottle virus firstly detected in Europe and other tobamoviruses

The tobamoviruses pose a recent threat to local varieties of Gernika pepper (cv. “Derio”) and Ibarra chili (cv. “Ibarroria”), causing a substantial decrease in both yield and fruit quality. Tobamoviruses such as Tomato mosaic virus (ToMV), Tobacco mild green mosaic virus (TMGMV) or Pepper mild mottle virus (PMMoV) are the viruses that cause the greatest impact in pepper crops of northern Spain. The emergence of a novel Tobamovirus, Chili pepper mild mottle virus (CPMMoV; GenBank MN164455.1), firstly identified in Europe, exacerbates these hazards. This situation has jeopardized the ongoing cultivation in some of the virus affected plots. A molecular marker-assisted backcrossing (MABC) breeding program was thus initiated to introduce the L3 and L4 resistance genes into these susceptible local varieties. MABC consisted of one crossing followed by four backcrosses and two self-fertilizations. After each crossbreeding event, the selection criteria of plants were based on plants resistance and fruits resemblance with the original sensitive varieties. This study allowed the homozygous introgression of resistance genes in the new obtained genotypes, reducing the reliance on hybrid seed production and associated costs. The resulting genotypes demonstrated robust resistance when inoculated with PMMoV 1.2, PMMoV 1.2.3, and CPMMoV. In addition, greenhouse yield trials were performed during two consecutive years to identify and select the most productive genotypes harboring the L3 or L4 gene, with the aim of registering some of them as commercial varieties. The productivity of the new selected genotypes did not display significant differences when compared to the original sensitive varieties. The introduction of genetic resistance in these local pepper varieties not only mitigates the immediate hazard posed by Tobamovirus but also fosters the preservation of local agricultural genetic resources, ensuring their continued cultivation and economic sustainability.

Comparison of Blast, Bacterial Leaf Blight and Brown Plant Hopper Introgressed Lines for Yield and Morphological Traits Over Original Line

Rice as a staple food for very large population suffers from various biotic and abiotic stresses. Among the biotic stresses, Blast, Bacterial leaf blight and Brown plant hopper (BPH) are considered to most potential threats that significantly affect the rice productivity. The use of chemical usage for controlling these diseases and pest attacks is not environmentally friendly and is expensive. Using Molecular Marker assisted backcross breeding program we have improved one of our hybrid parental line with 5 genes (xa13, Xa21, Pi54, Bph20 and Bph21). We have developed 15 near isogenic lines having similar agronomical characters as of recipient parent, they are promising for their direct induction in breeding program. Our results indicated that out of fifteen lines, Six lines i.e. GK 101-12, GK 101-15, GK 101-9, GK 101-5 and GK 101-2 out-performed in all the three locations tested, they had not only have very high level of resistance to BLB, Blast and BPH but also showed significantly higher yield compared to susceptible check as well as recipient parent. These lines may be used for improvement of our existing hybrid and also for developing of new hybrids with other CMS lines. These lines are also useful for developing new set of restorer lines through (R x R) and also useful for introgression of wide spectrum of resistance to the varieties/hybrid.

Evaluation for Parental Polymorphism and Identification of Microsatellites Linked to Drought Tolerance in Rice (Oryza sativa L.)

Aims: To identify the ready-to-use polymorphic microsatellite markers associated with drought tolerance for marker-assisted backcross breeding through a polymorphism survey between Jyothi and Chuvannamodan rice varieties.
 Place and Duration of Study: Centre for plant biotechnology and molecular biology, Thrissur, Kerala, India, during January to May 2023.
 Methodology: The genomic DNA of Jyothi and Chuvannamodan was isolated by following CTAB (Cetyltrimethyl Ammonium Bromide) method with modification. Isolated DNA from both varieties was subjected to PCR amplification using 208 Simple Sequence Repeats (SSRs) primer pairs distributed in 12 chromosomes. The amplified PCR products were electrophoresed in 3% agarose gel and separated fragments were visualized and documented in the Gel Documentation System. Different allele sizes produced by the same SSRs primers between two varieties are identified as polymorphic markers. Polymorphism per-cent was calculated, and frequency distribution and chromosome distribution of polymorphic markers were analyzed.
 Results: A total of 208 SSR primers were surveyed for the parental polymorphism. Out of which, 85 SSR primers exhibited clear polymorphism between Jyothi and Chuvannamodan and the remaining 123 were monomorphic primers. The amplicon size ranged from 83bp (RM430) to 495bp (RM18919) among the different primers. The survey revealed maximum parental polymorphism on chromosome 4 (69.23%), followed by chromosome 5 (64.28%), and minimum polymorphism on chromosome 8 (21.73%). The average per cent of polymorphism between the parents was 40.86%. Among the 85 polymorphic markers, 66 had dinucleotide repeats, 17 had trinucleotide repeats, and 1 had tetranucleotide repeats.
 Markers with dinucleotide repeats showed higher level of polymorphism. A group of 28 polymorphic markers were identified to be linked with traits including root-related traits, grain yield, leaf rolling and leaf drying under drought conditions.
 Conclusion: The polymorphic markers identified in the present study form the basis for tagging drought-tolerant QTLs/genes, fine mapping of those genes, and subsequently in marker-assisted breeding programs. The polymorphic markers linked with the QTLs for grain yield under drought can be pyramided in Jyothi through marker-assisted backcross breeding.

Improvement of bacterial blight resistance of the high yielding, fine-grain, rice variety, Gangavati sona through marker-assisted backcross breeding.

Gangavati sona (GS) is a high-yielding, fine-grain rice variety widely grown in the Tungabhadra command area in Karnataka, India; however, it is susceptible to bacterial blight (BB). Therefore, the present study was conducted to improve the GS variety for BB resistance. Three BB-resistant genes (xa5, xa13, and Xa21) were introgressed into the genetic background of susceptible cultivar GS through marker-assisted backcrossing (MABB) by using Improved samba Mahsuri (ISM), a popular, high-yielding, bacterial blight resistant rice variety as a donor parent. Foreground selection was carried out using gene-specific markers, viz., xa5FM (xa5), xa13prom (xa13), and pTA248 (Xa21), while background selection was carried out using well-distributed 64 polymorphic microsatellite markers. The true heterozygote F1 was used as the male parent for backcrossing with GS to obtain BC1F1. The process was repeated in BC1F1 generation, and a BC2F1 plant (IGS-5-11-5) possessing all three target genes along with maximum recurrent parent genome (RPG) recovery (86.7%) was selfed to obtain BC2F2s. At BC2F2, a single triple gene homozygote plant (IGS-5-11-5-33) with 92.6% RPG recovery was identified and advanced to BC2F5 by a pedigree method. At BC2F5, the seven best entries were selected, possessing all three resistance genes with high resistance levels against bacterial blight, yield level, and grain quality features equivalent to better than GS. The improved versions of GS will immensely benefit the farmers whose fields are endemic to BB.

Genetic Enhancement for Biotic Stress Resistance in Basmati Rice through Marker-Assisted Backcross Breeding.

Pusa Basmati 1509 (PB1509) is one of the major foreign-exchange-earning varieties of Basmati rice; it is semi-dwarf and early maturing with exceptional cooking quality and strong aroma. However, it is highly susceptible to various biotic stresses including bacterial blight and blast. Therefore, bacterial blight resistance genes, namely, xa13 + Xa21 and Xa38, and fungal blast resistance genes Pi9 + Pib and Pita were incorporated into the genetic background of recurrent parent (RP) PB1509 using donor parents, namely, Pusa Basmati 1718 (PB1718), Pusa 1927 (P1927), Pusa 1929 (P1929) and Tetep, respectively. Foreground selection was carried out with respective gene-linked markers, stringent phenotypic selection for recurrent parent phenotype, early generation background selection with Simple sequence repeat (SSR) markers, and background analysis at advanced generations with Rice Pan Genome Array comprising 80K SNPs. This has led to the development of Near isogenic lines (NILs), namely, Pusa 3037, Pusa 3054, Pusa 3060 and Pusa 3066 carrying genes xa13 + Xa21, Xa38, Pi9 + Pib and Pita with genomic similarity of 98.25%, 98.92%, 97.38% and 97.69%, respectively, as compared to the RP. Based on GGE-biplot analysis, Pusa 3037-1-44-3-164-20-249-2 carrying xa13 + Xa21, Pusa 3054-2-47-7-166-24-261-3 carrying Xa38, Pusa 3060-3-55-17-157-4-124-1 carrying Pi9 + Pib, and Pusa 3066-4-56-20-159-8-174-1 carrying Pita were identified to be relatively stable and better-performing individuals in the tested environments. Intercrossing between the best BC3F1s has led to the generation of Pusa 3122 (xa13 + Xa21 + Xa38), Pusa 3124 (Xa38 + Pi9 + Pib) and Pusa 3123 (Pi9 + Pib + Pita) with agronomy, grain and cooking quality parameters at par with PB1509. Cultivation of such improved varieties will help farmers reduce the cost of cultivation with decreased pesticide use and improve productivity with ensured safety to consumers.

Physiological and antioxidant responses associated with Sub1 gene introgressed rice (Oryza sativa L.) lines under complete submergence.

Rice is the only crop which is well adapted to aquatic environment but, it is unable to survive if completely submerged for several weeks. Breeding rice varieties with submergence tolerance is one of the best approaches to alleviate the adverse effect of submergence which requires the introgression of Sub1 gene into elite rice varieties. Hence, the study was undertaken to introgress submergence tolerant gene into the rice variety Jaya through Marker-Assisted Backcross Breeding. Also the physiological and biochemical responses like survival percentage, underwater shoot elongation, total carbohydrate content and superoxide dismutase activity were also studied in Sub1 introgressed lines. We could develop twenty Sub1 introgressed lines with Sub1 region of 3.1-5.1mb and with 80.0- 95.3% recurrent parent genome recovery. Sub1 introgressed Jaya lines and the tolerant checks FR13A and Swarna Sub1 had lower shoot elongation under water, higher superoxide dismutase activity (about 5 times) upto 4h after de-submergence which resulted in higher survival percentage. The reduced shoot elongation of tolerant varieties reduced the utilization of stored carbohydrate. Through our research we introgressed Sub1 gene into rice variety Jaya through Marker-Assisted Backcross Breeding and could study the physiological responses under submergence by which we confirmed the presence of Sub1 gene in these lines. These lines could be field evaluated and could be released as a new variety thus helping the farmers of flood prone areas of Kerala.

Development of dual stress-tolerant rice variety Jyothi with Saltol and Sub1 genes through marker-assisted backcross breeding

Development of dual stress-tolerant rice variety Jyothi with Saltol and Sub1 genes through marker-assisted backcross breeding

Marker-assisted enhancement of bacterial blight (Xanthomonas oryzae pv. oryzae) resistance in a salt-tolerant rice variety for sustaining rice production of tropical islands.

Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae is a major disease of rice, specially in the tropical regions of the world. Developing rice varieties with host resistance against the disease is the most effective and economical solution for managing the disease. Pyramiding resistance genes (Xa4, xa5, xa13,and Xa21) in popular rice varieties using marker-assisted backcross breeding (MABB) has been demonstrated as a cost-effective and sustainable approach for establishing durable BB resistance. Here, we report our successful efforts in introgressing four resistance genes (Xa4, xa5, xa13, and Xa21) from IRBB60 to CARI Dhan 5, a popular salt-tolerant variety developed from a somaclonal variant of Pokkali rice, through functional MABB. Both BB and coastal salinity are among the major challenges for rice production in tropical island and coastal ecosystems. Plants with four, three, and two gene pyramids were generated, which displayed high levels of resistance to the BB pathogen at the BC3F2 stage. Under controlled salinity microplot environments, the line 131-2-175-1223 identified with the presence of three gene pyramid (Xa21+xa13+xa5) displayed notable resistance across locations and years as well as exhibited a salinity tolerance comparable to the recurrent parent, CARI Dhan 5. Among two BB gene combinations (Xa21+xa13), two lines, 17-1-69-334 and 46-3-95-659, demonstrated resistance across locations and years, as well as salt tolerance and grain production comparable to CARI Dhan 5. Besides salinity tolerance, five lines, 17-1-69-179, 46-3-95-655, 131-2-190-1197, 131-2-175-1209, and 131-2-175-1239, exhibited complete resistance to BB disease. Following multilocation testing, potential lines have been identified that can serve as a prospective candidate for producing varieties for the tropical Andaman and Nicobar Islands and other coastal locations, which are prone to BB and coastal salinity stresses.

Pyramiding resistance genes for bacterial leaf blight (Xanthomonas oryzae pv. Oryzae) into the popular rice variety, Pratikshya through marker assisted backcrossing.

Bacterial leaf blight (BLB) is one of the major biotic stress in rice cultivation. Management techniques, such as the development of BLB-resistant cultivars, are required to lessen the severity of the disease attack and yield losses. Pratikshya was selected in the present investigation as the recipient parent, as it is one of the popular high-yielding rice varieties of Odisha, India, which is having excellent grain as well as cooking quality. However, Pratikshya is highly susceptible to BLB which is prevalent in Eastern Indian region. Three major BLB resistance genes xa5, xa13, and Xa21 from the donor source Swarna MAS (CR Dhan 800) were attempted to introduce into Pratikshya through a marker-assisted backcross breeding program. Those markers closely linked to the target genes were employed for foreground selection in the segregating generations till BC2F3. In each backcross generation, progenies containing all three targeted resistance genes and phenotypically more similar to the recipient parent, Pratikshya were selected and backcrossed. Screening of 1,598 plants of the BC2F2 population was conducted against BLB using Xoo inoculum and 35 resistant plants similar to Pratikshya were carried forward to the next generation. In the BC2F3 generation, 31 plants were found to possess all the three resistance genes. For background selection of plants carrying resistance genes 45 polymorphic SSR markers were employed. Evaluation of the pyramided lines at BC2F4 generation exhibited that, most pyramided lines were similar to Pratikshya in terms of morphological features and yield parameters, and some lines were superior to the recurrent parent in terms of morphological features and yield parameters. The three-gene pyramided lines showed a high level of resistance to BLB infection and are anticipated to offer a significant yield advantage over the recipient parent Pratikshya.The pyramided lines can further be used for multi-location trial, so as to be released as a variety or can be used as a potential donor for BLB resistance genes.

Discovering useful genetic variation in the seed parent gene pool for sorghum improvement.

Multi-parent populations contain valuable genetic material for dissecting complex, quantitative traits and provide a unique opportunity to capture multi-allelic variation compared to the biparental populations. A multi-parent advanced generation inter-cross (MAGIC) B-line (MBL) population composed of 708 F6 recombinant inbred lines (RILs), was recently developed from four diverse founders. These selected founders strategically represented the four most prevalent botanical races (kafir, guinea, durra, and caudatum) to capture a significant source of genetic variation to study the quantitative traits in grain sorghum [Sorghum bicolor (L.) Moench]. MBL was phenotyped at two field locations for seven yield-influencing traits: panicle type (PT), days to anthesis (DTA), plant height (PH), grain yield (GY), 1000-grain weight (TGW), tiller number per meter (TN) and yield per panicle (YPP). High phenotypic variation was observed for all the quantitative traits, with broad-sense heritabilities ranging from 0.34 (TN) to 0.84 (PH). The entire population was genotyped using Diversity Arrays Technology (DArTseq), and 8,800 single nucleotide polymorphisms (SNPs) were generated. A set of polymorphic, quality-filtered markers (3,751 SNPs) and phenotypic data were used for genome-wide association studies (GWAS). We identified 52 marker-trait associations (MTAs) for the seven traits using BLUPs generated from replicated plots in two locations. We also identified desirable allelic combinations based on the plant height loci (Dw1, Dw2, and Dw3), which influences yield related traits. Additionally, two novel MTAs were identified each on Chr1 and Chr7 for yield traits independent of dwarfing genes. We further performed a multi-variate adaptive shrinkage analysis and 15 MTAs with pleiotropic effect were identified. The five best performing MBL progenies were selected carrying desirable allelic combinations. Since the MBL population was designed to capture significant diversity for maintainer line (B-line) accessions, these progenies can serve as valuable resources to develop superior sorghum hybrids after validation of their general combining abilities via crossing with elite pollinators. Further, newly identified desirable allelic combinations can be used to enrich the maintainer germplasm lines through marker-assisted backcross breeding.

Introgression of recombinant 1RSWW.1BL without secalin and with Glu-B3/Gli-B1 into elite wheat cultivars through marker-assisted backcross breeding

Introgression of recombinant 1RSWW.1BL without secalin and with Glu-B3/Gli-B1 into elite wheat cultivars through marker-assisted backcross breeding

Evaluation of Powdery Mildew Resistance and Agronomic Performance of Pyramided Mungbean Lines from Marker- Assisted Selection

Gene pyramiding is an effective strategy to provide a durable and high level of powdery mildew (PM) resistance in mungbean [Vigna radiata (L.) R. Wilczek]. The objectives of this study were to evaluate PM resistance levels of pyramided BC2F1 progenies derived from marker-assisted selection of KING (recurrent parent) × SUPER5 (donor parent) cross and measure their agronomic performance under fi eld condition. The results showed that the progenies with marker combinations linked to both PM resistance genes including I85420 + I42PL222, and I27R565 were more resistant to PM (average disease severity score of 3.80) than KING, check cv. CN72, and the progenies without any marker combinations (scores of 8, 8 and 6.75, respectively). While the progenies carrying only one resistance gene were identifi ed as containing either I85420 + I42PL222, or I27R565 which showed intermediate response score of 5.83. In addition, most of the pyramided lines produced higher yield per plant than KING because of their superior pods per plant, clusters per plant, and seeds per pod. In the future, these pyramided lines with two PM resistance genes can be further developed into new PM resistant varieties via marker-assisted backcross breeding or be used as potential PM resistant sources in mungbean breeding programs.

Parental Polymorphism Survey for Identifying Genes/QTLs for BPH Resistance in Assam Land Race 10-3

The current research was taken up at Institute of Biotechnology (IBT), PJTSAU, Hyderabad Telangana, India in kharif from August to November, 2021 to access the parental polymorphism between 10-3 and Telangana Sona. 10-3, an Assam landrace, is a bold grain rice variety with high BPH resistance to biotype-4. Whereas, Telangana Sona (RNR 15048) is a short and slender grain variety of rice that is popular among farmers and consumers across Telangana and other Indian states due to its high yield and low glycemic index. Despite being blast resistant, Telangana Sona is susceptible to the insect pest Brown Plant hopper (BPH) Nilaparvata lugens (Stål). The present study aimed to characterize the parental polymorphism between 10–3 and Telangana sona to facilitate mapping of genetic loci linked with BPH resistance in the 10–3 and to optimize the retention of favourable alleles from Telangana sona during marker assisted backcross breeding. Eight hundred and sixteen highly variable Simple Sequence Repeats (SSRs) including thirty-five BPH linked markers covering 12 chromosomes were used to assess DNA polymorphism between 10-3 and Telangana Sona. Out of 816 markers screened, 97 (11.89%) were found to be polymorphic between 10–3 and Telangana Sona of which, 82 were genomic SSRs and 15 were BPH linked markers. Chromosome 6 exhibited highest polymorphism percentage with a value of 30%, indicating that this chromosome is useful in studying parental variation and lowest polymorphism percentage found on chromosome 5 (3.49%). These 97 polymorphic markers will help in construction of linkage map for identifying QTLs linked with BPH resistance and for marker assisted backcrossing breeding in Telangana Sona.

Parental polymorphism survey using rice microsatellite markers between VGD 1 and ADT 55 for marker-assisted backcross breeding program

Parental polymorphism survey using rice microsatellite markers between VGD 1 and ADT 55 for marker-assisted backcross breeding program

Stacking Multiple Genes Improves Resistance to Chilo suppressalis, Magnaporthe oryzae, and Nilaparvata lugens in Transgenic Rice.

The ability of various pests and diseases to adapt to a single plant resistance gene over time leads to loss of resistance in transgenic rice. Therefore, introduction of different pest and disease resistance genes is critical for successful cultivation of transgenic rice strains with broad-spectrum resistance to multiple pathogens. Here, we produced resistance rice lines with multiple, stacked resistance genes by stacking breeding and comprehensively evaluated their resistance to Chilo suppressalis (striped rice stemborer), Magnaporthe oryzae (rice blast), and Nilaparvata lugens (brown planthopper) in a pesticide-free environment. CRY1C and CRY2A are exogenous genes from Bacillus thuringiensis. Pib, Pikm, and Bph29 are natural genes in rice. CH121TJH was introduced into CRY 1C, Pib, Pikm, and Bph29. CH891TJH and R205XTJH were introduced into CRY 2A, Pib, Pikm, and Bph29. Compared with those observed in their recurrent parents, CH121TJH significantly increased the mortality of borers. The other two lines CH891TJH and R205XTJH are the same result. Three lines introduction of Pib and Pikm significantly reduced the area of rice blast lesions, and introduction of Bph29 significantly reduced seedling mortality from N. lugens. Introduction of the exogenous genes had relatively few effects on agronomic and yield traits of the original parents. These findings suggest that stacking of rice resistance genes through molecular marker-assisted backcross breeding can confer broad spectrum and multiple resistance in differently genetic backgrounds.