水稻内卷叶突变体acl3的鉴定及调控基因的功能分析

doi:10.3724/SP.J.1006.2025.42052

摘要/Abstract

摘要：

适度卷叶有利于异交结实和提高作物群体光合效率，在杂交水稻制种及生产中具有重要的应用价值。我们从甲基磺酸乙酯EMS诱变体库中鉴定到1个稳定遗传的内卷叶突变体acl3 (adaxially curled leaf 3)，该突变体表现出株高和结实率降低、千粒重显著提高。石蜡切片表明，叶片上表皮维管束间的泡状细胞数量减少和面积变小是导致acl3极度内卷的主要原因。遗传分析表明，acl3的突变性状受1对单隐性核基因调控，初步定位在第7染色体Indel标记07g89和07g498之间409 kb的物理范围内。对突变体acl3及其野生型西大1B进行全基因组测序，利用IGV软件查找acl3定位区间的序列差异，发现注释基因LOC_Os07g01240/ SRL1/CLD1在第6外显子上存在1个G到A的碱基替换，初步确定为候选基因。通过构建互补载体转化突变体acl3，表型鉴定结果显示互补植株的卷叶、株高、籽粒大小等突变性状均恢复至野生型水平，表明ACL3是SRL1的一个新等位基因，并具有“一因多效”的特性。qRT-PCR分析结果显示，卷叶相关基因REL2和NAL7极显著上调；进一步分析生长素(IAA)途径相关基因的表达变化发现，生长素合成基因YUCCA2、应答基因ARF1和ARF7、原初响应基因IAA10、IAA21、IAA22等均有不同程度的上调，这表明ACL3可能通过生长素途径调控水稻株型的发育过程。

关键词: 水稻(Oryza sativa L.), 泡状细胞, 卷叶, SRL1, 生长素

Abstract:

Moderate leaf curling is beneficial for cross-pollination, fruit set, and enhancing the photosynthetic efficiency of crop canopies, making it highly valuable for hybrid rice seed production and cultivation. To elucidate the molecular mechanisms underlying leaf rolling in rice, we identified a stably inherited adaxial leaf-rolling mutant, acl3 (adaxially curled leaf 3), from an ethyl methanesulfonate (EMS)-mutagenized population. In addition to leaf curling, acl3 exhibits reduced plant height and seed-setting rate, along with a significant increase in thousand-grain weight. Histological analysis using paraffin sections revealed that the extreme curling in acl3 primarily results from a reduction in both the number and area of bulliform cells between vascular bundles in the adaxial epidermis. Genetic analysis indicated that the acl3 mutant phenotype is controlled by a single recessive nuclear gene, which was preliminarily mapped to a 409 kb physical region between Indel markers 07g89 and 07g498 on chromosome 7. Whole-genome sequencing of acl3 and the wild-type Xida 1B was performed, and sequence variations within the acl3 mapping interval were identified using IGV software. A G-to-A base substitution was detected in the sixth exon of the annotated gene LOC_Os07g01240/SRL1/CLD1, leading to a structural change in the encoded protein. This gene was preliminarily identified as the candidate gene for ACL3. Complementary vector construction and transformation of the acl3 mutant restored the mutant traits—leaf curling, plant height, and grain size—to wild-type levels, confirming that ACL3 is a new allele of SRL1 with pleiotropic effects. Quantitative real-time PCR (qRT-PCR) analysis revealed significant upregulation of leaf-rolling-related genes REL2 and NAL7. Further investigation of auxin IAA pathway-related genes showed that auxin synthesis gene YUCCA2, auxin response genes ARF1 and ARF7, and primary auxin response genes IAA10, IAA21, and IAA22 were all upregulated to varying degrees, suggesting that ACL3 may regulate rice plant architecture development through the auxin signaling pathway.

Key words: rice (Oryza sativa L.), bulliform cells, curled leaf, SRL1, auxin

雷松翰, 范骏扬, 车艳奕, 代永东, 郑雨萌, 田维江, 桑贤春, 王晓雯. 水稻内卷叶突变体acl3的鉴定及调控基因的功能分析[J]. 作物学报, 0, (): 0-.

LEI Song-Han, FAN Jun-Yang, CHE Yan-Yi, DAI Yong-Dong, ZHENG Yu-Meng, TIAN Wei-Jiang, SANG Xian-Chun, WANG Xiao-Wen. Identification of an adaxially-curled-leaf mutant acl3 and function analysis of the regulated gene in rice (Oryza sativa L.)[J]. Acta Agronomica Sinica, 0, (): 0-.

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