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Not AvailableThe realization of yield potential of rice (Oryza sativa L.) hybrids developed through the “three-line system” largely depends on genetic purity of parental lines. Cytoplasmic male sterile (CMS) lines often get contaminated with cognate isonuclear maintainer lines during multiplication. We report identification of a rice mitochondrial DNA sequence, possessing a repeat motif that is polymorphic between the wild abortive (WA)-CMS lines and their cognate isonuclear maintainer lines. We have designed a primer pair flanking the unique repeat and two fragments of different size were amplified in all WA-CMS and their maintainer lines through PCR. The PCR assay was able to precisely detect the impurities in a commercial seed lot of a popular CMS line, IR58025A. The possibility of utilizing this marker as a replacement for the morphology-based (grow-out test) GOT in accurate detection of contaminants in CMS seed stocks is discussed.Not Availabl

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Not AvailableHost plant resistance is the preferred management strategy for Asian rice gall midge (Orseolia oryzae), a serious pest in many rice-growing countries. Identification of simple sequence repeat (SSR) markers that are tightly linked to pest resistance genes can accelerate development of gene pyramids for durable/multiple resistance. Based on conventional and molecular allelism tests, we report herein that rice genotype Aganni possesses Gm8 gene, conferring hypersensitive independent (HR– type) resistance to gall midge biotypes GMB1, GMB2, GMB3, GMB4, and GMB4M. The gene Gm8 was mapped to chromosome 8 within a 400-kbp region, and the SSR markers RM22685 and RM22709 flank the gene closely. Using these closely linked flanking markers, nine other gall midge-resistant genotypes were identified as carrying the same gene Gm8. Through marker-assisted selection, Gm8 has been introgressed into an elite bacterial blight-resistant cultivar, Improved Samba-Mahsuri (IS).Not Availabl

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Not AvailableThe intensity of rice false smut disease in selected states of northwest and south India was studied. In northern Indian states as a whole, disease incidence (percentage of false smut-infected tillers) varied from 2% to 75%. In the state of Haryana, maximum infection was recorded on hybrids like PA 6444 and PA 6129 while in Punjab state, 10–20% disease incidence was recorded in popular inbred rice varieties like PR 114, PA 116 and PAU 201. In the southern state of Tamil Nadu, the disease incidence varied from 5% to 85%. A heavy incidence of the disease was noticed in variety BPT 5204 and due to this, the air above the infected field gave a black smoky appearance from a distance as a result of release of spore mass in the atmosphere. In severe cases the number of infected grains reached even more than 100 per panicle. The pathogen Ustilaginoidea virens was isolated in potato dextrose agar medium and was characterized by both pathogenicity test and molecular analysis. Under glasshouse conditions, when a conidial suspension of the pathogen was injected during boot leaf stage of the rice variety TN1, typical smut balls were observed. The identity of the pathogen was further confirmed through polymerase chain reaction (PCR) analysis using U. virens-specific internal transcribed spacer (ITS) primers. The primer pair US 1-5/US3-3 and US2-5/US4-3 amplified 380 bp and 232 bp product, respectively, which are typical for the U. virens fungus.Not Availabl

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Not AvailableThe recessive rice gall midge resistance gene, gm3 identified in the rice breeding line RP2068-18-3-5 confers resistance against five of the seven Indian biotypes of the Asian rice gall midge Orseolia oryzae. We report here tagging and fine-mapping of gm3 gene, development of a functional marker for it and demonstrated its use in marker-assisted selection (MAS). A mapping population consisting of 302 F10 recombinant inbred lines derived from the cross TN1 (susceptible)/RP2068-18-3-5, was screened against gall midge biotype 4 (GMB4) and analyzed with a set of 89 polymorphic SSR markers distributed uniformly across the rice genome. Two SSR markers, RM17480 and gm3SSR4, located on chromosome 4L displayed high degree of co-segregation with the trait phenotype and flanked the gene. In silico analysis of the genomic region spanning these two markers contained 62 putatively expressed genes, including a gene encoding an NB-ARC (NBS-LRR) domain containing protein. A fragment of this gene was amplified with the designed marker, NBcloning 0.9 Kb from the two susceptible TN1, Improved Samba Mahsuri (B95-1) and two resistant cultivars, RP 2068-18-3-5 and Phalguna (with Gm2 gene). The amplicons were observed to be polymorphic between the susceptible and resistant genotypes and hence were cloned and sequenced. A new primer, gm3del3, which was designed based on sequence polymorphism, amplified fragments with distinct size polymorphism among RP2068-18-3-5, Phalguna and TN1 and B95-1 and displayed no recombination in the entire mapping population. Expression of the candidate NB-ARC gene in the susceptible TN1 and the resistant RP2068-18-3-5 plants following infestation with GMB4 was analyzed, through real-time reverse transcription PCR. Results showed twofold enhanced expression in RP2068-18-3-5 plants, but not in TN1 plants, 120 h after infestation. Amino acid sequence and structure analysis of the proteins coded by different alleles of gm3 gene showed deletion of eight amino acids due to an early stop codon in RP2068-18-3-5 resulting in a change in the functional domain of the protein. The gm3del3 was used as a functional marker for introgression of gm3 gene into the genetic background of the elite bacterial blight resistant cultivar Improved Samba Mahsuri (B95-1) through MAS.Not Availabl

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Not AvailableIn changing climate scenario, heat stress caused by increased atmospheric CO2 is a major concern for rice productivity. There is a need to decipher the mechanisms of heat stress susceptibility and tolerance response of rice cultivars considering that high temperature is detrimental to growth and development of rice crop. The present study was designed to understand the heat stress response in heat-susceptible (Vandana) and heat-tolerant (N22) cultivars of rice. Rice seedlings were subjected to short-duration (24 h, SDS) and long-duration (5 days, LDS) heat stress (42 °C/36 °C, day/night). Besides the heat stress, recovery response (REC) of both the cultivars was also studied. Physiological parameters (chlorophyll content and membrane thermostability) and root/shoot length analysis revealed that N22 has better efficiency in recovering from heat stress. In particular, root tissue of N22 showed increased thermotolerance during SDS and LDS when compared with Vandana. In addition to physiological studies, gene expression pattern of 13 genes including heat shock transcription factors and heat shock proteins and 9 microRNAs (miRNAs) was analyzed in root and shoot of both the genotypes during various treatments. Gene and miRNA expression studies showed that root tissue of N22 was more responsive during SDS and LDS, suggesting important function of roots in heat stress tolerance. Further, during recovery, root tissue of both the genotypes showed more significant change in gene expression than shoot which signifies the vital role of plant root system in heat stress recovery response. Very high expression of an unknown iron-sulfur cluster-binding protein OsFd involved in electron transport activity was observed in root tissue of N22 during all the stress treatments. This study shows that better recovery and efficient root system play an important role in heat tolerance trait of N22.Not Availabl

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Not AvailableThe cultivation of rice (Oryza sativa L.), a major food crop, requires ample water (30 % of the fresh water available worldwide), and its productivity is greatly affected by drought, the most significant environmental factor. Much research has focussed on identifying quantitative trait loci, stress-regulated genes and transcription factors that will contribute towards the development of climate-resilient/tolerant crop plants in general and rice in particular. The transcription factor DREB1A, identified from the model plant Arabidopsis thaliana, has been reported to enhance stress tolerance against drought stress. We developed transgenic rice plants with AtDREB1A in the background of indica rice cultivar Samba Mahsuri through Agrobacterium-mediated transformation. The AtDREB1A gene was stably inherited and expressed in T1 and T2 plants and in subsequent generations, as indicated by the results of PCR, Southern blot and RT-PCR analyses. Expression of AtDREB1A was induced by drought stress in transgenic rice lines, which were highly tolerant to severe water deficit stress in both the vegetative and reproductive stages without affecting their morphological or agronomic traits. The physiological studies revealed that the expression of AtDREB1A was associated with an increased accumulation of the osmotic substance proline, maintenance of chlorophyll, increased relative water content and decreased ion leakage under drought stress. Most of the homozygous lines were highly tolerant to drought stress and showed significantly a higher grain yield and spikelet fertility relative to the nontransgenic control plants under both stressed and unstressed conditions. The improvement in drought stress tolerance in combination with agronomic traits is very essential in high premium indica rice cultivars, such as Samba Mahsuri, so that farmers can benefit in times of seasonal droughts and water scarcity.Not Availabl

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Not AvailableIn this study, complete genome of a south Indian isolate of Rice tungro spherical virus (RTSV) from Andhra Pradesh (AP) was sequenced, and the predicted amino acid sequence was analysed. The RTSV RNA genome consists of 12,171 nt without the poly(A) tail, encoding a putative typical polyprotein of 3,470 amino acids. Furthermore, cleavage sites and sequence motifs of the polyprotein were predicted. Multiple alignment with other RTSV isolates showed a nucleotide sequence identity of 95 % to east Indian isolates and 90 % to Philippines isolates. A phylogenetic tree based on complete genome sequence showed that Indian isolates clustered together, while Vt6 and PhilA isolates of Philippines formed two separate clusters. Twelve recombination events were detected in RNA genome of RTSV using the Recombination Detection Program version 3. Recombination analysis suggested significant role of 5′ end and central region of genome in virus evolution. Further, AP and Odisha isolates appeared as important RTSV isolates involved in diversification of this virus in India through recombination phenomenon. The new addition of complete genome of first south Indian isolate provided an opportunity to establish the molecular evolution of RTSV through recombination analysis and phylogenetic relationship.Not Availabl

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Not AvailableRice tungro disease, one of the major constraints for rice production in South and Southeast Asia, is caused by a combination of two viruses: Rice tungro spherical virus (RTSV) and Rice tungro bacilliform virus (RTBV). The disease is characterized by typical symptoms produced in rice plants; however, reliance on symptoms for the detection of disease and presence of viral particles is not always correct. The present study was undertaken to utilize the polymerase chain reaction (PCR) and RT-PCR techniques to diagnose the RTBV and RTSV particles, respectively. Primers were designed from highly conserved regions of viral genomes to amplify the ORF-I and ORF-IV regions of RTBV and CP3 genetic region of RTSV. The detection method was standardized and validated using tungro-infected leaf samples collected from Cuttack (Odisha), Puducherry UT, Coimbatore, Kanyakumari (TN), Patancheru and Rajendranagar (Andhra Pradesh). Insect vectors (green leaf hopper, Nephotettix spp.) were collected from virus infected plants maintained in glass house. All the three primers used in PCR and RTPCR were found reliable in detecting the virus particles in insects also.Not Availabl

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Not AvailableRice blast is a devastating fungal disease, which limits rice production severely. To identify novel alleles of blast resistant gene, Pi54, and to understand its diversity exist among wild Oryza species and landraces sequence-based allele mining strategy was employed. In this study, sixteen Pi54 alleles were sequenced from landraces and wild Oryza species which have genome ranging from AA to EE. Overall analysis confirmed that the alleles derived from wild species had more divergence than the landraces. Among all wild Oryza alleles, Pi54 aus derived from O. australiensis, showed the highest diversity. Comparison of protein domains revealed that LRR region had more variations than NBS region. Haplo-groups among the ecotypes were analyzed based on their Pi54 sequence. Interestingly, four different haplo-groups were identified, that were supported by phylogeny analysis. Among 16 analyzed alleles, two alleles, Pi54 ab, Pi54 btj derived from landraces-Amana Bavo and Boha Thulasi Joha showed superior phenotypic reaction than the reference allele (Pi54 Tetep), and these alleles were validated by allelism test. These alleles have potential application in blast resistance breeding programs and identified SNPs and Indel among the alleles can be useful in development of allele specific markers. The present study helped to gain insights into the evolutionary adaptation of the resistant gene and its allelic distribution and diversity among the Oryza species.Department of Biotechnology, Government of Indi