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鉴定水稻在热应激条件下差异表达的基因。

Identification of differentially expressed genes under heat stress conditions in rice (Oryza sativa L.).

机构信息

Department of Genetics and Plant Breeding, S.V. Agricultural College, Acharya NG Ranga Agricultural University (ANGRAU), Tirupati, AP, 517502, India.

Department of Plant Breeding, Institute of Frontier Technology, RARS, Acharya NG Ranga Agricultural University (ANGRAU), Tirupati, AP, 517502, India.

出版信息

Mol Biol Rep. 2020 Mar;47(3):1935-1948. doi: 10.1007/s11033-020-05291-z. Epub 2020 Feb 17.

Abstract

Rice production in recent years is highly affected by rapidly increasing temperatures in the tropical and sub-tropical countries, which threatens the sustainable production in near future. Hence, understanding the heat tolerance mechanism and evolving tolerant varieties is an immense need in the staple crop rice. An experiment has been conducted to identify differentially expressed genes in rice under heat stress conditions by employing a diverse set of 32 rice genotypes that includes reported heat tolerant genotypes Nagina 22 (N22) and Dular. Screening of the genotypes at field conditions during Summer-2018 for reproductive stage heat tolerance (wherein the mean minimum (29.8 °C) and maximum (38.4 °C) temperatures surpassed optimum temperatures (25 °C night/30 °C day) required for rice flowering and grain filling stages) and lab conditions employing thermal induction response (TIR) technique to know the genotype's acquired thermal tolerance revealed that the genotype FR13A (indica landrace) showed highest overall performance for multitude of traits viz., 95.29% of spikelet fertility (SF-%) at field level and 100% seedling survival percentage (SSP) at sub-lethal temperatures under laboratory conditions. The relative performance (under TIR) across all the three traits viz., relative shoot length (RSL) (4.91), relative root length (RRL) (equal to the control) and relative seedling dry weight (RSDW) (6.94) over control is high when compared to the other genotypes under study. However, the highly susceptible genotype PUSA1121 performed with 43.59 of SF%, 73.33% SSP, - 43.59 of RSL, - 36.02 of RRL over control. Hence, these contrasting genotypes were used for molecular analysis for identification of differentially expressed genes by employing 29 heat related gene specific primers. Five genes viz., OsGSK1, TT1, HSP70-OsEnS-45, OsHSP74.8 and OsHSP70 have shown differential expression between the two genotypes. Hence, the genotype FR13A, an 'indica' genotype, can be utilized in heat tolerance breeding programmes as donor parent in addition to the reported 'aus' genotypes, N22 and Dular. To our knowledge this is the first indica genotype identified for heat tolerance. The HSP70s, TT1 and OsGSK1 that proved with differential expression might be used for identification of gene specific InDels and thereby to develop functional markers that help in the marker assisted introgression breeding to develop heat tolerant varieties that can sustain production under dramatically changing climatic conditions.

摘要

近年来,热带和亚热带国家的气温迅速上升,严重影响了水稻的产量,这对未来的可持续生产构成了威胁。因此,了解水稻的耐热机制和培育耐热品种是水稻这一主要粮食作物的迫切需求。

本研究通过对包括耐热基因型 Nagina 22 (N22) 和 Dular 在内的 32 个水稻基因型进行实验,以鉴定水稻在热胁迫条件下差异表达的基因。在 2018 年夏季的田间条件下对基因型进行筛选,以评估其生殖期耐热性(平均最低(29.8°C)和最高(38.4°C)温度超过了水稻开花和灌浆阶段所需的最适温度(25°C 夜间/30°C 白天)),并采用热诱导反应(TIR)技术在实验室条件下评估基因型的获得性耐热性。结果表明,基因型 FR13A(籼稻地方品种)在多个性状方面表现出最佳整体性能,即田间水平的小穗育性(SF-%)为 95.29%,实验室亚致死温度下的种子活力百分比(SSP)为 100%。与其他研究中的基因型相比,三个性状(相对茎长(RSL)(4.91)、相对根长(RRL)(等于对照)和相对幼苗干重(RSDW)(6.94))的相对性能(在 TIR 下)均较高。然而,高度敏感的基因型 PUSA1121 的 SF-%、SSP、RSL 和 RRL 分别为 43.59、73.33、-43.59 和-36.02。因此,这些对比鲜明的基因型被用于分子分析,通过使用 29 个与热相关的基因特异性引物来鉴定差异表达的基因。发现两个基因型之间有五个基因(OsGSK1、TT1、HSP70-OsEnS-45、OsHSP74.8 和 OsHSP70)表达存在差异。因此,基因型 FR13A(籼稻基因型)可以作为耐热育种计划的供体亲本加以利用,除了已报道的耐热基因型 N22 和 Dular 外,还可以利用它。据我们所知,这是第一个被鉴定为耐热的籼稻基因型。已证明具有差异表达的 HSP70s、TT1 和 OsGSK1 可能用于鉴定基因特异性 InDels,并由此开发功能标记,以帮助进行基于标记的渐渗导入育种,从而培育出能在气候剧烈变化条件下持续生产的耐热品种。

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