Sarla Neelamraju

Characterization of a Nagina22 rice mutant for heat tolerance and mapping of yield traits

BackgroundHeat is one of the major factors that considerably limit rice production. Nagina 22 (N22) is a deep-rooted, drought and heat tolerant aus rice cultivar. This study reports the characterization of a previously isolated dark green leaf mutant N22-H-dgl219 (NH219) which showed reduced accumulation of reactive oxygen species in leaf under 40°C heat conditions.The mutant was characterized for several traits in field under ambient (38°C) and heat stress (44°C) conditions by raising temperature artificially from flowering stage till maturity by covering plants with polythene sheets during dry season 2011. Yield traits were mapped in 70 F2 segregants of IR64 × NH219 and 36 F2 segregants of its reciprocal cross.ResultsLeaf proteome analysis using two-dimensional gel electrophoresis from N22 and NH219 showed distinct constitutive expression of ribulose bisphosphate carboxylase large chain precursor (EC 4.1.1.39) in NH219 under ambient growth condition. Heat stress resulted in reduction of all 11 traits except plant height in both N22 and NH219. The extent of reduction was more in N22 than in NH219. Both pollen viability and spikelet fertility were not reduced significantly in N22 and NH219 but reduced by 20% in IR64.ConclusionNH219 is more tolerant to heat stress than wild type N22 as its percent yield reduction is lesser than N22. Single marker analysis showed significant association of RM1089 with number of tillers and yield per plant, RM423 with leaf senescence, RM584 with leaf width and RM229 with yield per plant.

Cytokinin delays dark-induced senescence in rice by maintaining the chlorophyll cycle and photosynthetic complexes

Highlight Cytokinin induces functional stay-greenness by maintaining the Chl a/b ratios and the stability of photosynthetic complexes during dark-induced senescence in rice.

Os11Gsk gene from a wild rice, Oryza rufipogon improves yield in rice

Chromosomal segments from wild rice species Oryza rufipogon, introgressed into an elite indica rice restorer line (KMR3) using molecular markers, resulted in significant increase in yield. Here we report the transcriptome analysis of flag leaves and fully emerged young panicles of one of the high yielding introgression lines IL50-7 in comparison to KMR3. A 66-fold upregulated gene Os11Gsk, which showed no transcript in KMR3 was highly expressed in O. rufipogon and IL50-7. A 5-kb genomic region including Os11Gsk and its flanking regions could be PCR amplified only from IL50-7, O. rufipogon, japonica varieties of rice-Nipponbare and Kitaake but not from the indica varieties, KMR3 and Taichung Native-1. Three sister lines of IL50-7 yielding higher than KMR3 showed presence of Os11Gsk, whereas the gene was absent in three other ILs from the same cross having lower yield than KMR3, indicating an association of the presence of Os11Gsk with high yield. Southern analysis showed additional bands in the genomic DNA of O. rufipogon and IL50-7 with Os11Gsk probe. Genomic sequence analysis of ten highly co-expressed differentially regulated genes revealed that two upregulated genes in IL50-7 were derived from O. rufipogon and most of the downregulated genes were either from KMR3 or common to KMR3, IL50-7, and O. rufipogon. Thus, we show that Os11Gsk is a wild rice-derived gene introduced in KMR3 background and increases yield either by regulating expression of functional genes sharing homology with it or by causing epigenetic modifications in the introgression line.

Expression patterns of QTL based and other candidate genes in Madhukar × Swarna RILs with contrasting levels of iron and zinc in unpolished rice grains

BACKGROUND Identifying QTLs/genes for iron and zinc in rice grains can help in biofortification programs. Genome wide mapping showed 14 QTLs for iron and zinc concentration in unpolished rice grains of F7 RILs derived from Madhukar × Swarna. One line (HL) with high Fe and Zn and one line (LL) with low Fe and Zn in unpolished rice were compared with each other for gene expression using qPCR. 7 day old seedlings were grown in Fe+ and Fe- medium for 10 days and RNA extracted from roots and shoots to determine the response of 15 genes in Fe- conditions. RESULTS HL showed higher upregulation than LL in shoots but LL showed higher upregulation than HL in roots. YSL2 was upregulated only in HL roots and YSL15 only in HL shoots and both up to 60 fold under Fe- condition. IRT2 and DMAS1 were upregulated 100 fold and NAS2 1000 fold in HL shoot. NAS2, IRT1, IRT2 and DMAS1 were upregulated 40 to 100 fold in LL roots. OsZIP8, OsNAS3, OsYSL1 and OsNRAMP1 which underlie major Fe QTL showed clear allelic differences between HL and LL for markers flanking QTL. The presence of iron increasing QTL allele in HL was clearly correlated with high expression of the underlying gene. OsZIP8 and OsNAS3 which were within major QTL with increasing effect from Madhukar were 8 fold and 4 fold more expressed in HL shoot than in LL shoot. OsNAS1, OsNAS2, OsNAS3, OsYSL2 and OsYSL15 showed 1.5 to 2.5 fold upregulation in flag leaf of HL when compared with flag leaf of Swarna. CONCLUSION HL and LL differed in root length, Fe concentration and expression of several genes under Fe deficiency. The major distinguishing genes were NAS2, IRT2, DMAS1, and YSL15 in shoot and NAS2, IRT1, IRT2, YSL2, and ZIP8 in roots. The presence of iron increasing QTL allele in HL at marker locus close to genes also increased upregulation in HL.

Transgressive segregation for yield traits in Oryza sativa IR58025B X Oryza meridionalis Ng. Bc2F3population under irrigated and aerobic conditions

Wild species of the genus Oryza are a good source of beneficial alleles for enhancing rice yield under normal and adverse conditions. BC2F3 population was derived from a cross between Oryza sativa IR58025B and Oryza meridionalis Ng. (2n = 24, AA) a heat tolerant wild species to evaluate 12 yield traits under irrigated and aerobic conditions. Analysis of variance and genetic estimates indicated there is substantial genetic variation among progenies under both conditions. Grain yield had high heritability (61.9%) and genetic advance (36.4%) under irrigated conditions but moderate heritability (49.6%) and genetic advance (13.3%) under aerobic conditions indicating that selection for yield will be effective under both conditions. Panicle number, grain number, spikelet fertility, and test weight showed significant positive correlation with grain yield under both conditions. Families out-performing IR58025B for yield under both conditions were obtained providing evidence that phenotypically inferior O. meridionalis contributed to yield increase. This species can be a novel source of natural genetic variation for the improvement of rice under irrigated as well as under aerobic condition.

Quantitative trait loci and candidate genes for yield and related traits in Madhukar x Swarna RIL population of rice

Yield of popular rice varieties such as Swarna grown in rainfed lowlands and Madhukar grown in flood prone areas needs to be continuously improved. Recombinant inbred lines (RILs) were developed from the cross between two indica cultivars Madhukar and Swarna. QTLs were mapped using 110 markers in 168 RILs. In all, 26 QTLs were mapped for yield and five related traits on chromosomes 1, 2, 3, 6, 7, 8, 10, 11, and 12. QTL for plant height and days to flowering were co-located between RM23147 — RM337 on chromosome 8. RM251, RM314, and RM1135 were significantly associated with plant height and OsYSL17 was significantly linked with grain yield. Epistatic interaction was detected for plant height and number of tillers. Several candidate genes reported for yield and related traits underlie the QTL regions.

Genotype × Environment interactions of Nagina22 rice mutants for yield traits under low phosphorus, water limited and normal irrigated conditions

Multi environment testing helps identify stable genotypes especially for adverse abiotic stress situations. In the era of climate change and multiple abiotic stresses, it becomes important to analyze stability of rice lines under both irrigated and stress conditions. Mutants are an important genetic resource which can help in revealing the basis of natural variation. We analyzed 300 EMS induced mutants of aus rice cultivar Nagina22 (N22) for their G × E interaction and stability under low phosphorus (P), water limited and irrigated conditions. Environmental effect and interaction were more significant than genotypic contribution on grain yield (GY), productive tillers (TN) and plant height (PH) under these three environmental conditions in dry season, 2010. GY and TN were more affected by low P stress than by water limited condition, but PH was not significantly different under these two stresses. Mutants G17, G209, G29, G91, G63 and G32 were stable for GY in decreasing order of stability across the three environments but G254 and G50 were stable only in low P, G17 and G45 only in water limited and G295 and G289 only in normal irrigated condition. We then selected and evaluated 3 high yielding mutants, 3 low yielding mutants and N22 for their stability and adaptability to these 3 environments in both wet and dry seasons for six years (2010–2015). The most stable lines based on the combined analysis of 12 seasons were G125 (NH210) under normal condition, G17 (NH686), G176 (NH363) and G284 (NH162) in low P condition and G176 (NH363) under water limited condition. G176 was the best considering all 3 conditions. When screened for 15 Pup1 gene-specific markers, G176 showed alleles similar to N22. While two other low-P tolerant lines G17 and G65 showed N22 similar alleles only at k-1 and k-5 but a different allele or null allele at 13 other loci. These stable mutants are a valuable resource for varietal development and to discover genes for tolerance to multiple abiotic stresses.

Down regulation of a heavy metal transporter gene influences several domestication traits and grain Fe-Zn content in rice

Biofortification of rice (Oryza sativa L.) would alleviate iron and zinc deficiencies in the target populations. We identified two alleles 261 and 284 of a Gramineae-specific heavy metal transporter gene OsHMA7 by analyzing expression patterns and sequences of genes within QTLs for high Fe & Zn, in Madhukar x Swarna recombinant inbred lines (RILs) with high (HL) or low (LL) grain Fe & Zn. Overexpression of 261 allele increased grain Fe and Zn but most of the transgenic plants either did not survive or did not yield enough seeds and could not be further characterized. Knocking down expression of OsHMA7 by RNAi silencing of endogenous gene resulted in plants with altered domestication traits such as plant height, tiller number, panicle size and architecture, grain color, shape, size, grain shattering, heading date and increased sensitivity to Fe and Zn deficiency. However, overexpression of 284 allele resulted in transgenic lines with either high grain Fe & Zn content (HL-ox) and tolerance to Fe and Zn deficiency or low grain Fe & Zn content (LL-ox) and phenotype similar to RNAi-lines. OsHMA7 transcript levels were five-fold higher in the HL-ox plants whereas LL-ox and RNAi plants showed 2-3 fold reduced levels compared to Kitaake control. Spraying LL-ox and RNAi lines with Fe & Zn at grain filling stage resulted in increased grain yield, significant increase in Fe & Zn content and brown pericarp. Altered expression of OsHMA7 influenced transcript levels of iron-responsive genes indicating cellular Fe-Zn homeostasis and also several domestication-related genes in rice. Our study shows that a novel heavy metal transporter gene influences yield and grain Fe & Zn content and has potential to improve rice production and biofortification.