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Transcriptomic and Genetic Analysis of Seed Germination Vigor under Low Temperature in Weedy Rice WR04-6

ABSTRACT: Low temperature is one of the major factors affecting rice germination, and low tempera-ture germination (LTG) is an important agronomic trait. Although genetic variation is abundant in rice germplasm resources, the molecular mechanism of LTG remains poorly understood. In this study, we first proved that weedy rice WR04-6 had significantly better low-temperature germination (LTG) ability at 10°C than the cultivated rice Qishanzhan (QSZ). RNA-seq was used to investigate the gene expression of WR04-6 and QSZ at 10, 12 and 14 days of seed germination at 10°C. The results of GO enrichment and KEGG en-richment revealed that the differentially expressed genes between WR04-6 and QSZ were mainly concentrated on the response to starch catabolic processes and the response to ab-scisic acid. This is consistent with the results of α-amylase activity, ABA and GA treat-ment. A recombinant inbred line (RIL) population derived from a cross between WR04-6 and QSZ and its high density SNP genetic map were used to detect quantitative trait loci (QTL) for low temperature germination rates at 10°C for 14 days. The results showed that two new QTLs were located on chromosome 3 and chromosome 12. Combined with the mapped QTLs and RNA-seq differential genes (DEGs), sixteen candidate genes potentially associated with LTG were identified. Validation of expression of the candidates by qRT-PCR were consistent with the RNA-seq data. These results will enable us to under-stand the genetic basis of LTG in weedy rice and provide new genetic resources for gener-ation of rice germplasm with LTG.

ORGANISM(S): Oryza sativa

PROVIDER: GSE222929 | GEO | 2023/03/01

REPOSITORIES: GEO

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Project description:Background: Weedy rice (Oryza sativa L.) is a worldwide problem in rice production, being highly tolerant to sub-optimal nutrient levels hence competitive in nutrient acquisition. To understand the function of genes that are potentially involved in the high nutrient acquisition ability of weedy rice, we compared the transcriptomes of strawhull weedy red rice (tolerant to N deficit) with the rice cultivar ‘Wells’ (intolerant to N deficit), by examining profiles in flag leaves at panicle initiation under low and optimum N levels. Strawhull weedy red rice and cultivar ‘Wells’ were grown in nutrient solution with NH4NO3 concentration manipulated to simulate optimum and deficient N conditions. Changes in gene expression in leaf tissues were analyzed at three conditions: N deficiency, and at 24- and 48-h NH4NO3 supplementation after N starvation. Differential gene expression on weedy red rice was evaluated using oligonucleotide arrays representing 44,974 rice gene models. Overall, comparative real-time PCR analysis of 21 candidate genes identified from the microarray data between weedy red rice and cultivar ‘Wells’ supported our hypothesis that key genes involved in N assimilation are expressed differentially at N- deficient conditions between the tolerant and intolerant strains. Results: Eight candidate genes showed significant differences in expression at one of the time points: N and starch metabolism-related [alanine aminotransferase (OsAlaAT) locus ID Os10g25140.1; soluble starch synthase 2-1(OsSSSII1), Os10g30156.1; and soluble starch synthase 2-3(OsSSSII3), Os06g12450.1]; cell structure-related [alpha-L-fucosidase 2 precursor (OsFUCA2), Os06g06250.1]; signal transduction [two-component response regulator-like(OsPRR1), Os02g40510.1 and EF hand family protein (OsPOLC2_JUNOX), Os02g50060.1]; and transcription factors [zinc finger, C2H2 type family protein(OsC2H2Znf), Os11g06840.1 and Myb-like DNA-binding domain (OsMYB), Os01g62660.1]. Genome-wide gene expression analysis of weedy rice showed that nitrite reductase (Os01g25484.1; Os01g25484.2; Os01g25484.3) was most highly induced at N starvation and was most deeply repressed at 24 h of N-stress recovery. A few other genes, namely SANT/MYB (Os01g47370.1), chaperonin (Os02g54060.1; Os02g54060.2), protein phosphatase (Os09g15670.1), polyamine transporter (Os01g61044.1), trehalose-6-phosphate synthase (Os02g54820.2), uracil phosphoribosyltransferase (Os05g38170.1), an MIKCc type-box transcription factor (Os02g52340.1), a cell homeostasis-related uncharacterized protein (Os02g16880.1), a protease inhibitor (Os07g18990.1), dehydrin (Os11g26760.1), and cytochrome P450 (Os11g05380.1) were also strong indicators of starvation and recovery. Conclusions: Weedy rice has N-stress adaptive mechanisms that are probably distinct to the mechanisms in most cultivars. This mechanism potentially contributes to its high vigor and competitive advantage over most rice cultivars under sub-optimal nutrient levels. Expression of key genes involved in nitrate assimilation, trehalose synthesis, and protein modification appeared to be critical for adaptation to N stress in weedy rice. N-stress tolerance of weedy red rice appeared to be due at least in part to the ability to sustain C fixation and starch synthesis during N starvation.

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Transcriptomic and Genetic Analysis of Seed Germination Vigor under Low Temperature in Weedy Rice WR04-6

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