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1St Indian Rice Congress 1st Indian Rice Congress - 2020 Rice Research and Development for Achieving Sustainable Development Goals Theme - I : Enhancing rice productivity and quality IRC/TM-1/PP-86 IDENTIFICATION OF GENOMIC REGIONS LINKED TO DORMANCY RELATED TRAITS USING BULK SEGREGANT ANALYSIS IN RICE (ORYZA SATIVA L.) D. B. K. V. Mani*, B. Krishna Veni, V. Roja and S. N. Umar Department of Genetics and Plant Breeding, Agricultural College, Bapatla-522101, Andhra Pradesh, India *Corresponding authors email: [email protected] A total of 119 F6:7 recombinant inbred lines of be associated with dormancy. Out of the 4 polymorphic a cross between BPT 2231 (non-dormant parent) and markers used in the present study, RM346 is notified MTU 1001 (dormant parent) was analyzed to identify as a dormancy linked marker from previous studies. the markers associated with dormancy. Parental The other 3 markers viz., RM22565, RM7051 and polymorphism survey with 188 SSR markers revealed RM10793 identified as dormancy linked markers in 10 polymorphic markers between the parents. The bulk the present study needs further validation on alternative segregant analysis results revealed that four markers set of population or a set of germplasm lines for their showed polymorphism between these bulks. The further utilization in the marker assisted breeding association of putative markers viz., RM346, programme. Based on germination percentage, RM22565, RM7051 and RM10793 identified based physiological parameters and genotyping studies, the on DNA pooling from selected segregant was analyzed lines SD 3, SD 12, SD 15 and SD 68 may be used in by Single Marker Analysis (SMA).The results of SMA future breeding programme as donor parents for seed revealed that RM22565 on chromosome 8 showed dormancy. significant association with germination (%) at 5 days Keywords: SSR markers, Bulk segregant after harvesting indicating that the chromosomal region analysis, Single marker analysis linking to marker RM 22565 on chromosome 8 may December 8-9, 2020 | ICAR-NRRI, Cuttack 753006, Odisha, India 205 1st Indian Rice Congress - 2020 Rice Research and Development for Achieving Sustainable Development Goals Theme - I : Enhancing rice productivity and quality IRC/TM-1/PP-87 MAPPING OF QTLS FOR ZINC DEFICIENCY TOLERANCE IN RIL POPULATION OF RICE (ORYZA SATIVA L.) Bapsila Loitongbam1*, P. K. Singh2, R. P. Sah3, Rabin Sahu3, Priyanka Irungbam4 and Ng. T. Chanu4 1College of Agriculture, Central Agricultural University (Imphal), Pasighat-791102, Arunachal Pradesh 2Institute of Agricultural Science, Banaras Hindu University, Varanasi-221102, Uttar Pradesh,India 3Division of Crop Improvement, ICAR-National Rice Research Institute, Cuttack-753006, Odisha, India 4College of Horticulture and Forestry, Central Agricultural University (Imphal), Pasighat-791102, Arunachal Pradesh,India *Corresponding author: [email protected] Rice is one of the important cereal crops determine the response of RIL population with regard consuming more than half of the world’s population to Zn deficiency tolerance under Zn deficient soil and 95% of it is being produced and consumed in Asia. conditions to identify and map QTLs associated with it Zinc (Zn) deficiency is one of the major abiotic stresses using SSR markers. limiting rice growth and its production. Among the OBJECTIVES cereals, rice is considered as highly sensitive to Zn 1. Estimation of zinc concentration in RIL deficiency and it is the most important micronutrient, mapping population. restricted the rice growth and its yield. Zn deficiency in 2. Construction of genetic linkage map and rice plants causes various symptoms that usually appear identification of genomic regions having QTL for zinc 2 to 3 weeks after transplanting (WAT) rice seedlings. deficiency tolerance. Zn deficiency leads to develop brown blotches and streaks in leaves that may ultimately fuse to cover older METHODOLOGY leaves entirely, plants remain stunted and in severe cases The experimental materials consisted of 236 may die. Plants with severe Zn deficiency undergo RILs (F7) mapping population developed from a cross symptoms like leaf bronzing with stunting growth and between Kinandang Patong and A69-1. The field ultimately leads to low grain yield. Zn deficiency is screening under Zn deficiency was carried out at two associated with a diverse range of soil conditions with locations viz., BHU, Varanasi, India and Acharya high pH (>7.0) alkaline soil, low availability of Zn Narendra Deva University of Agriculture & Technology content, prolonged submergence in water (Neue and (NDUAT), Ayodhya. The experiment was laid out in Lantin, 1994). Use of RIL populations for mapping of an alpha lattice design with two replications at both the QTLs is considered to be highly advantageous due to locations. The RILs mapping population and their the facts that multiple selfing events increase the parents were evaluated for traits associated with zinc recombination events which permit a finer mapping of deficiency viz., leaf bronzing score, plant mortality, root/ QTLs. Once the RILs are established having fixed shoot zinc concentration for each replications at both genotypes as homozygote, these lines are often the locations and their means were used for the statistical repeatedly used for investigating QTLs of diverse analysis in a Zn deficient field condition. The leaf phenotypes under different environments provided the bronzing (LB) was scored visually five times at 3 WAT parents involved were contrasting for the trait of and at late vegetative stage through 1-9 scales, along interest. Therefore, the aim of this study was to with parents and two sensitive checks. The total 206 December 8-9, 2020 | ICAR-NRRI, Cuttack 753006, Odisha, India 1st Indian Rice Congress - 2020 Rice Research and Development for Achieving Sustainable Development Goals Theme - I : Enhancing rice productivity and quality genomic DNA was isolated from the leaf of twenty- and one minor QTLs at Ayodhya for Zn deficiency one days old seedling using the method described by tolerance with Logarithm of odd (LOD) threshold value Doyle and Doyle, (1990) with some modifications. higher than 3 were identified. A total of six QTLs at DNA of the two parents, Kinandang Patong (sensitive Varanasi and five QTLs at Ayodhya by CIM methods to Zn deficiency) and A69-1(tolerant to Zn deficiency) governing leaf bronzing, plant mortality, root Zn was extracted for polymorphism detection using 697 concentration and shoot Zn concentration were number of SSR markers. 81 SSR markers were detected across two locations. These QTLs were polymorphic with the parents and used for genotyping located mostly in chromosome 2, 4 and 6. the 236 RILs mapping population. Linkage maps were Leaf bronzing scored (1-9) was conducted five constructed using MAPMAKER/EXP version 3.0b times at seedling to late vegetative stage in 236 RILs (Lander et al., 2009) following Kosambi Function along with parents and two sensitive checks, IR-74 (Kosambi, 1944). Linkage between the markers and and Dular. Among the 236 RILs, 33 lines were highly the QTL was detected by a statistical test called the tolerant at, Varanasi and 35 lines at, Ayodhya showing Logarithm of Odds (LOD) score method. QTLs were no symptoms of Zn deficiency and healthy growth with detected by Composite Interval Mapping (CIM) leaf bronzing score of 1 (Growth and tillering nearly procedure of Windows QTL Cartographer v.2.5 normal; healthy). While 17lines and 21 lines which software (Wang et al., 2006). scored (9) were almost dead or dying at , Varanasi RESULTS and , Ayodhya respectively. The results of the analysis of variance CONCLUSION (ANOVA) for, Varanasi and, Ayodhya revealed The present study could identify major QTLs significant variation among the RILs for all the on chromosome 4 for all the zinc deficiency tolerant associated traits with Zn deficiency (P<0.01). Out of traits under studied in both the locations and one major 697 SSR markers, 81 markers showed parental QTL on chromosome 6 at BHU location. The identified polymorphism however, only 74 markers were QTLs were found to be novel and introgression of these included in constructing a linkage map and 7showed new QTLs into elite rice breeding lines will be very unlinked of the remaining markers to the linkage map. promising. About 11.62% of primers were exhibited parental REFERENCE polymorphism. The linkage map covered 1353.3 cM Wissuwa, M., Ismail, A.M., Yanagihara, S., 2006. distance employing Kosambi mapping function, resulting Effects of zinc deficiency on rice growth and in an average marker interval of 18.28 cM. QTL genetic factors contributing to tolerance. Plant analysis was performed using WinQtlCart 2.5 for Physiology 142(2), 731-741. identification of QTLs related to Zn deficiency Xu, Y., Zhu, L., Xiao, J., Huang, N., McCouch, S.R., tolerance. The association of molecular markers and 1997. Chromosomal regions associated with phenotypic data was performed using Composite segregation distortion of molecular markers in interval mapping analysis with 3 as LOD threshold for F2 backcross double haploid and recombinant detecting a QTL. A total of five [major (>10 R2) and inbred population in rice (Oryza sativa L.). one minor (<10 R2)] QTLs at Varanasi and four major Molecular Genetics and Genomics 253, 535- 545 December 8-9, 2020 | ICAR-NRRI, Cuttack 753006, Odisha, India 207 1st Indian Rice Congress - 2020 Rice Research and Development for Achieving Sustainable Development Goals Theme - I : Enhancing rice productivity and quality IRC/TM-1/PP-88
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