Effect analysis of S5-interacting genes on rice hybrid sterility using nontransgenic gamete killer

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science Pub Date : 2024-12-13 DOI:10.1016/j.plantsci.2024.112357

Jie Li , Fu Huang , Yingxia Jiang, Jianglei Rao, Yourong Fan, Jiangyi Yang

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引用次数: 0

Abstract

While hybrids between japonica and indica rice exhibit strong heterosis, they often suffer from hybrid sterility (HS). Hybrid fertility of the embryo sac is predominantly regulated by a three-gene system (comprising closely linked ORF3, ORF4 and ORF5) at rice S5 locus. The cooperation of ORF5+ and ORF4+ can result in endoplasmic reticulum (ER) stress and sporophytically kill all embryo sacs, while ORF3+ can gametophytically protect the residing embryo sac. We previously identified four S5-interacting genes (SIGs) using a transgenic line BL^ORF5⁺ (Balilla carrying transgenic ORF5+) and a wide compatibility variety Dular (DL or D). Homozygote and hemizygote of ORF5+ transgene had significantly different spikelet fertility (SF), which disturbed the phenotypic effects of SIGs. However, HS effects of SIGs under the endogenous (nontransgenic) gamete killer remained unknown. We formerly constructed a semisterile near isogenic line (NIL) S5-BL/NJ by introgressing S5 fragment of indica rice Nanjing11 (NJ or N, carrying ORF3+ORF4-ORF5+ haplotypes) into the genome of japonica rice Balilla (BL or B, carrying ORF3-ORF4+ORF5- haplotypes). The gamete-protecting effect of ORF3+ in NJ may confuse SF effect of the SIGs, so we knocked out ORF3+ of S5-NJ/NJ and crossed it with BL to get gamete-killing S5-BL/NJ^ΔORF3+, which can kill all (KA) gametes (abbreviated as enS5KA). Compared with the exS5KA line (a NIL carrying ORF5+ transgenic, wihch can kill all gamete), the enS5KA line conferred SIGs a more pronounced SF effect. The enS5KA,SIG-DDDD (four SIGs carry homozygous DL alleles) genotype caused a SF of about 78 %, while SF of exS5KA,SIG-DDDD was only about 62 %. Moreover, all SIGs acted in a sporophytic manner without segregation distortion of genotype. Although enS5KA,SIG-DDDD plants had high SF, the ER stress still existed. The ovule section revealed that enS5KA,SIG-BBBB genotype (four SIGs carry homozygous BL allele, with ER stress and SF < 5 %) caused abnormal degradation of nucellar cells and functional megaspores. In contrast, enS5KA,SIG-DDDD genotype preserved most nucellar cells and functional megaspores. These results lay the foundation for further research on HS mechanism of S5 and SIGs and cloning of candidate genes.

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利用非转基因配子杀手分析 S5 相互作用基因对水稻杂种不育性的影响。

虽然粳稻和籼稻的杂交种表现出很强的异交性，但它们经常会出现杂交不育症（HS）。胚囊的杂交育性主要受水稻 S5 基因座上的三基因系统（包括紧密相连的 ORF3、ORF4 和 ORF5）调控。ORF5+ 和 ORF4+ 的协同作用会导致内质网（ER）应激，孢子体会杀死所有胚囊，而 ORF3+ 则能配子体保护驻留的胚囊。此前，我们利用转基因品系 BLORF5+（携带转基因 ORF5+ 的 Balilla）和广相容性品种 Dular（DL 或 D）鉴定出了四个 S5 相互作用基因（SIG）。ORF5+转基因的同源杂合子和半杂合子的小穗生育力（SF）明显不同，这干扰了SIGs的表型效应。然而，SIGs 在内源（非转基因）配子杀手下的 HS 效应仍然未知。以前，我们通过将籼稻南京11（NJ或N，携带ORF3+ORF4-ORF5+单倍型）的S5片段导入粳稻Balilla（BL或B，携带ORF3-ORF4+ORF5-单倍型）的基因组，构建了半固定近等基因系（NIL）S5-BL/NJ。NJ中ORF3+的配子保护作用可能会混淆SIG的SF效应，因此我们敲除了S5-NJ/NJ的ORF3+，并将其与BL杂交，得到了能杀死所有（KA）配子的配子杀伤型S5-BL/NJΔORF3+（简称enS5KA）。与exS5KA品系（携带ORF5+的转基因NIL，可杀死所有配子）相比，enS5KA品系赋予SIG的SF效应更明显。enS5KA,SIG-DDDD（四个 SIG 携带同源 DL 等位基因）基因型的 SF 值约为 78%，而 exS5KA,SIG-DDDD 的 SF 值仅为 62%。此外，所有 SIG 都以孢子体方式发挥作用，基因型没有发生分离失真。虽然 enS5KA,SIG-DDDD 植株的 SF 值很高，但仍然存在 ER 胁迫。胚珠切片显示，enS5KA,SIG-BBBB 基因型（4 个 SIG 携带同源 BL 等位基因，具有 ER 胁迫，SF < 5%）导致核细胞异常退化和功能性巨孢子。相反，enS5KA,SIG-DDDD 基因型保留了大部分核细胞和功能性巨孢子。这些结果为进一步研究 S5 和 SIG 的 HS 机制以及克隆候选基因奠定了基础。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Plant Science 生物-生化与分子生物学

CiteScore

9.10

自引率

1.90%

发文量

322

审稿时长

33 days

期刊介绍： Plant Science will publish in the minimum of time, research manuscripts as well as commissioned reviews and commentaries recommended by its referees in all areas of experimental plant biology with emphasis in the broad areas of genomics, proteomics, biochemistry (including enzymology), physiology, cell biology, development, genetics, functional plant breeding, systems biology and the interaction of plants with the environment. Manuscripts for full consideration should be written concisely and essentially as a final report. The main criterion for publication is that the manuscript must contain original and significant insights that lead to a better understanding of fundamental plant biology. Papers centering on plant cell culture should be of interest to a wide audience and methods employed result in a substantial improvement over existing established techniques and approaches. Methods papers are welcome only when the technique(s) described is novel or provides a major advancement of established protocols.