Introgression of bacterial leaf blight (BLB) resistant gene, Xa7 into MARDI elite variety, MR219 by marker assisted backcrossing (MABC) approach

Bacterial leaf blight (BLB) is one of the major rice diseases in Malaysia. This disease causes substantial yield loss as high as 70%. Development of rice varieties which inherited BLB resistant traits is a crucial approach to promote and sustain rice industry in Malaysia. Hence, this study aims were to enhance BLB disease resistant characters of high yielding commercial variety MR219 through backcross breeding approach with supporting tool of marker-assisted selection (MAS). Broad spectrum BLB resistance gene, Xa7 from donor parent IRBB7 were introgressed into the susceptible MR219 (recurrent parent) using two flanking markers ID7 and ID15. At BC3F4, we managed to generate 19 introgressed lines with homozygous Xa7 gene and showed resistant characteristics as donor parent when it was challenged with Xanthomonas oryzae pv. oryzae through artificial inoculation. Recurrent parent MR219 and control variety, MR263 were found to be severely infected by the disease. The improved lines exhibited similar morphological and yield performance characters as to the elite variety, MR219. Two lines, PB-2-107 and PB-2-34 were chosen to be potential lines because of their outstanding performances compared to parent, MR219. This study demonstrates a success story of MAS application in development of improved disease resistance lines of rice against BLB disease.

Development of an SNP Marker Set for Marker-Assisted Backcrossing Using Genotyping-By-Sequencing in Tetraploid Perilla

High-quality molecular markers are essential for marker-assisted selection to accelerate breeding progress. Compared with diploid species, recently diverged polyploid crop species tend to have highly similar homeologous subgenomes, which is expected to limit the development of broadly applicable locus-specific single-nucleotide polymorphism (SNP) assays. Furthermore, it is particularly challenging to make genome-wide marker sets for species that lack a reference genome. Here, we report the development of a genome-wide set of kompetitive allele specific PCR (KASP) markers for marker-assisted recurrent selection (MARS) in the tetraploid minor crop perilla. To find locus-specific SNP markers across the perilla genome, we used genotyping-by-sequencing (GBS) to construct linkage maps of two F2 populations. The two resulting high-resolution linkage maps comprised 2,326 and 2,454 SNP markers that spanned a total genetic distance of 2,133 cM across 16 linkage groups and 2,169 cM across 21 linkage groups, respectively. We then obtained a final genetic map consisting of 22 linkage groups with 1,123 common markers from the two genetic maps. We selected 96 genome-wide markers for MARS and confirmed the accuracy of markers in the two F2 populations using a high-throughput Fluidigm system. We confirmed that 91.8% of the SNP genotyping results from the Fluidigm assay were the same as the results obtained through GBS. These results provide a foundation for marker-assisted backcrossing and the development of new varieties of perilla.

Improvement of the submergence stress tolerance of local South Sumatran rice through the introgression of the Sub1 gene by using marker-assisted selection

Submergence stress due to unpredictable soil flooding is one of the main constraints encountered in rainfed growing areas, especially in Southern Sumatran riparian swamps. The development of submergence-stress-tolerant cultivars through the introgression of Sub1 via marker-assisted backcrossing (MABC) is an ideal solution. This study was carried out during 2020 at Sriwijaya University, Palembang, Indonesia, with the aim to select Sub1-introgressed lines in BC3F1 generations on the basis of MABC and to screen out the SSR markers that were unlinked to the target gene for application in subsequent background selection studies. Results revealed that almost all the backcrossed progenies segregated from the rice parental cultivars ‘FR13A’ and ‘Pegagan’. The backcrossed lines showed significantly improved submergence stress tolerance and recovery rates compared with their parents. Sub1 introgression into the BC3F1 generation was confirmed by the tightly linked Sub1 marker SUB1C173, and marker RM23915 was used for recombinant selection. These markers followed the expected marker segregation ratio in accordance with the Mendelian single gene model. In the parental polymorphism survey, 84 out of 237 SSR markers that were unlinked to the target loci were found to be available for background study. Twenty-seven backcrossed lines were selected on the basis of foreground selection. Seven plants were selected on the basis of the recombinant marker RM23915. Five backcrossed lines were further selected on the basis of submergence stress tolerance and agronomic performance.

Drought Tolerance and Application of Marker-Assisted Selection in Sorghum

Sorghum is an important staple food crop in drought prone areas of Sub-Saharan Africa, which is characterized by erratic rainfall with poor distribution. Sorghum is a drought-tolerant crop by nature with reasonable yield compared to other cereal crops, but such abiotic stress adversely affects the productivity. Some sorghum varieties maintain green functional leaves under post-anthesis drought stress referred to as stay-green, which makes it an important crop for food and nutritional security. Notwithstanding, it is difficult to maintain consistency of tolerance over time due to climate change, which is caused by human activities. Drought in sorghum is addressed by several approaches, for instance, breeding drought-tolerant sorghum using conventional and molecular technologies. The challenge with conventional methods is that they depend on phenotyping stay-green, which is complex in sorghum, as it is constituted by multiple genes and environmental effects. Marker assisted selection, which involves the use of DNA molecular markers to map QTL associated with stay-green, has been useful to supplement stay-green improvement in sorghum. It involves QTL mapping associated with the stay-green trait for introgression into the senescent sorghum varieties through marker-assisted backcrossing by comparing with phenotypic field data. Therefore, this review discusses mechanisms of drought tolerance in sorghum focusing on physiological, morphological, and biochemical traits. In addition, the review discusses the application of marker-assisted selection techniques, including marker-assisted backcrossing, QTL mapping, and QTL pyramiding for addressing post-flowering drought in sorghum.

Translational Chickpea Genomics Consortium to Accelerate Genetic Gains in Chickpea (Cicer arietinum L.)

The Translational Chickpea Genomics Consortium (TCGC) was set up to increase the production and productivity of chickpea (Cicer arietinum L.). It represents research institutes from six major chickpea growing states (Madhya Pradesh, Maharashtra, Andhra Pradesh, Telangana, Karnataka and Uttar Pradesh) of India. The TCGC team has been engaged in deploying modern genomics approaches in breeding and popularizing improved varieties in farmers’ fields across the states. Using marker-assisted backcrossing, introgression lines with enhanced drought tolerance and fusarium wilt resistance have been developed in the genetic background of 10 elite varieties of chickpea. Multi-location evaluation of 100 improved lines (70 desi and 30 kabuli) during 2016–2017 and 2018–2019 enabled the identification of top performing desi and kabuli lines. In total, 909 Farmer Participatory Varietal Selection trials were conducted in 158 villages in 16 districts of the five states, during 2017–2018, 2018–2019, and 2019–2020, involving 16 improved varieties. New molecular breeding lines developed in different genetic backgrounds are potential candidates for national trials under the ICAR-All India Coordinated Research Project on Chickpea. The comprehensive efforts of TCGC resulted in the development and adoption of high-yielding varieties that will increase chickpea productivity and the profitability of chickpea growing farmers.

Disease resistance of improved MR220 lines against Pyricularia oryzae Cavara and their preliminary agronomic performance

Blast disease caused by Pyricularia oryzae is one of the most destructive fungal diseases of rice in Malaysia. Utilisation of resistant varieties is the most efficient management approach towards reducing yield losses. The line IRTP21683 with the Pi9 gene has shown strong resistance against the isolate MPO988.3 of pathotype P_0.0, the most prevalent P. oryzae pathotype in Malaysia. Crossing of IRTP21683 was undertaken with the recurrent parent MR220, a susceptible elite Malaysian rice variety, using a marker assisted backcrossing technique with two simple sequence repeat markers, RM19776 and RM7311, as the tag for the Pi9 gene. Twenty BC₃F₄ lines with the Pi9 gene were resistant when challenged with MPO 988.3. The cluster analysis based on seven agronomic parameters showed that the resistant BC₃F₄ lines could be divided into four groups, of which the members in group 1 and 2 have shown comparable or better performance than MR220. Five lines in group 1, B220PI9-3-48, B220PI9-3-76, B220PI9-3-77, B220PI9-3-79 and B220PI9-3-82 showed outstanding yield performance with early maturation.

Introgression of QTLs for Drought Tolerance into Farmers’ Preferred Sorghum Varieties

Sorghum is a major staple food crop for the people in semi-arid areas of Africa and Asia. Post-flowering drought is a global constraint of sorghum production. The study aimed to improve stay-green (STG) characteristics of farmer-preferred sorghum varieties in Tanzania using marker-assisted backcrossing. A total of 752 individuals representing five BC2F1 populations and their parents were genotyped using previously reported KASP markers linked with STG 3A and STG 3B quantitative trait loci (QTL). In the BC2F1 populations, the maximum number of individuals with heterozygous alleles were observed in S35*Pato background (37) whereas only seven individuals derived from the B35*Wahi parents’ background contained heterozygous alleles. Of the 30 single nucleotide polymorphism (SNP) markers, favourable alleles were observed at 18 loci in BC2F1 populations. In the BC2F1 generation, the highest (0.127 kg/panicle) grain yield was observed in the B35*NACO Mtama 1 background population. The genotypic analysis revealed the presence of favourable alleles in homozygous conditions at markers loci associated with STG 3A and STG 3B QTLs in BC2F3 populations, suggesting successful introgression of STG QTLs from the donor parents to the recurrent parents. Across water irrigation regimes, the highest (0.068 kg/panicle) mean grain weight was observed in the genotype NA316C. Therefore, our study demonstrated the utility of marker-assisted backcrossing for drought tolerance improvement of locally adapted sorghum varieties in Africa.

Construction of high-density genetic map and QTL mapping in Nicotiana tabacum backcrossing BC4F3 population using whole-genome sequencing

Backcrossing is a powerful tool for plant breeding. The improved marker-assisted backcrossing intends to transfer targeted genes or quantitative trait loci (QTLs) of interest from a donor parent into a recurrent parent. In this study, a tobacco BC4F3 population was generated using Y3 and K326 as hybrid parents and YF1-1 as F₁ parents. High-throughput sequencing data of 381 pedigree populations were used to construct high-density genetic maps containing 24 142 high-quality single nucleotide polymorphism (SNP) markers with an average genetic distance of 0.59 cM. A genome module analysis was then performed for all the offspring. A total of forty-three candidate QTLs for six agronomics traits were identified. This study provides original biomarkers for tobacco breeding and offers clues for prospective backcrossing applications in other plants.