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作物学报

作物学报 ›› 2023, Vol. 49 ›› Issue (6): 1699-1707.doi: 10.3724/SP.J.1006.2023.22028

• 研究简报 • 上一篇    下一篇

水稻籽粒伸长突变体lgdp的鉴定与基因定位

林孝欣(), 黄明江, 韦祎, 朱洪慧, 王子怡, 李忠成, 庄慧, 李彦羲, 李云峰*(), 陈锐*()   

  1. 西南大学水稻研究所/西南大学农业科学研究院/转基因植物与安全控制重庆市重点实验室, 重庆 400715
  • 收稿日期:2022-05-09 接受日期:2022-10-10 出版日期:2023-06-12 网络出版日期:2022-11-17
  • 通讯作者: *陈锐, E-mail: chenruin998@gmail.com;李云峰, E-mail: liyf1980@swu.edu.cn
  • 作者简介:E-mail: 2393551889@qq.com
  • 基金资助:
    国家自然科学基金项目(32172044)

Identification and gene mapping of long grain and degenerated palea (lgdp) in rice (Oryza sativa L.)

LIN Xiao-Xin(), HUANG Ming-Jiang, WEI Yi, ZHU Hong-Hui, WANG Zi-Yi, LI Zhong-Cheng, ZHUANG Hui, LI Yan-Xi, LI Yun-Feng*(), CHEN Rui*()   

  1. Rice Research Institute, Southwest University/Academy of Agricultural Sciences, Southwest University/Transgenic Plants and Safety Control, Chongqing Key Laboratory, Chongqing 400715, China
  • Received:2022-05-09 Accepted:2022-10-10 Published:2023-06-12 Published online:2022-11-17
  • Contact: *E-mail: chenruin998@gmail.com;E-mail: liyf1980@swu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(32172044)

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摘要：

水稻粒形与产量和营养品质密切相关, 挖掘水稻粒形发育相关基因并解析其分子机制, 对提高水稻产量、改善籽粒营养品质具有重要意义。利用甲基磺酸乙酯(ethyl methanesulfonate, EMS)处理籼稻品种西大1B, 获得了1个水稻粒形突变体, 命名为long grain and degenerated palea (lgdp)。lgdp表现出外稃伸长, 从而导致籽粒长度增加的特征, 进一步扫描电镜分析发现, lgdp籽粒变长主要原因是其外稃细胞数目极显著增加。遗传分析表明该性状受1对隐性基因调控; 利用lgdp与ZH11杂交构建的F2分离群体, 通过BSA法将目标基因定位在3号染色体分子标记ZLN43和ZLN-1之间, 物理距离大约810 kb。通过转录测序和PCR分析初步确认LGDP候选基因编码一个MADS-box基因。qPCR分析表明, LGDP可能通过负向调控GW7/GL7、GS3、TGW6等水稻粒长正向调控因子的表达, 从而影响了颖壳细胞数目的增殖, 进而影响籽粒长度。本研究结果为应用LGDP基因改良水稻粒形提供了新的资源。

关键词: 水稻, 粒形, 细胞数目, 基因定位

Abstract:

The grain shape, which consists of grain length and grain width, is the primary determinant of grain yield and one of the important appearance quality traits in rice. It is of great significance to identify the related genes associated with grain shape and to study molecular mechanisms for improving the yield and quality of rice. In this study, a long grain mutant named long grain and degenerated palea (lgdp) deriving from EMS (ethyl methane sulfonate) mutation groups of Xida 1B was reported. In lgdp mutant, the elongation of lemma resulted in a long grain. Further SEM analysis revealed that the main reason for lemma elongation was the extremely significant increase in the number of glume cells. Genetic analysis showed that the lgdp trait was regulated by a pair of recessive genes. Using BSA method and the F2 population crossing lgdp with ZH11, the target gene was located between the molecular markers ZLN43 and ZLN-1 on chromosome 3, with a physical distance of about 810 kb. The analysis of RNA-seq and PCR indicated the LGDP candidate gene might encode a MADS-box protein. The qPCR referred that LGDP negatively regulated the relative expression levels of several positive grain length regulatory factors, GW7/GL7, GS3, TGW6, which affected the cell proliferation of glumes and the grain length. The results of this study laid a foundation for the molecular function analysis of LGDP gene in the future.

Key words: rice (Oryza sativa L.), grain shape, cell number, gene mapping

图 1

野生型和突变体lgdp的表型观察 A: 野生型小穗; B: 图A中小穗去掉外稃后的形态; C: 野生型小穗的横切面; D: 图C的局部放大; E: 野生型早期幼穗原基; F, K: lgdp的Type1和Type2两种小穗类型; G: 图F中小穗去除外稃后的形态; H: lgdp小穗的横切面; I: 图H的局部放大; J: Type1早期幼穗原基; L: 图K中小穗去除外稃后的形态: M: lgdp小穗的横切面; N: 图M的局部放大; O: Type2早期幼穗原基。缩写: le: 外稃; sl: 护颖; lo: 浆片: pa: 内稃; pi: 雌蕊。标尺: A为22 mm; B为20 mm; C为240 μm; D为75 μm; E, J, O为100 μm; F, G, K, L为23 mm; H为300 μm; I为78 μm; M为300 μm, N为80 μm。"

图2

野生型(WT)和突变体lgdp的株型与粒形 A: 野生型和lgdp的植株; B: 野生型和lgdp籽粒宽度比较; C: 野生型和lgdp籽粒(去壳)宽度比较; D: 野生型和lgdp籽粒长度比较; E: 野生型和lgdp籽粒(去壳)长度比较; F: 株高统计; G: 节间长度统计; H: 穗长统计; I: 一次枝梗数统计; J: 二次枝梗数统计; K: 每穗小花数; L: 每穗实粒数统计; M: 粒宽统计; N: 籽粒(去壳)宽度统计; O: 粒长统计; P: 籽粒(去壳)长度统计; Q: 结实率统计; R: 千粒重统计。**表示在0.01概率水平差异显著, *表示在0.05概率水平差异显著。标尺: A为40 cm; B为33 mm; C为40 mm; D为90 mm。"

图3

野生型(WT)和突变体lgdp颖壳的扫描电镜(SEM)观察 A: 野生型的成熟籽粒颖壳; B: lgdp突变体的成熟籽粒颖壳; C, D: 扫描电镜观察外稃外表皮; E, F: 扫描电镜观察内稃外表皮; G: 外稃长度统计; H: 内桴长度统计; I: 外稃外表皮纵行细胞数目统计; J: 内稃外表皮纵行细胞数目统计: K: 外稃外表皮细胞大小统计; L: 内稃外表皮细胞大小统计。缩写: le: 外稃; pa: 内稃。**表示在0.01概率水平差异显著, *表示在0.05概率水平差异显著。标尺: A、B为5 mm; C~F为300 μm。"

表1

野生型和lgdp突变体F2分离的卡方测验"

名称
Name		 父本
Male parent		 母本
Female parent		 野生型
Wild type		 突变型
Mutant type		 卡方值
Chi-square value (χ20.05=3.84)
组合1 Combination 1 lgdp 56S 355 106 0.89
组合2 Combination 2 lgdp ZH11 176 44 2.67
组合3 Combination 3 lgdp NIP 70 13 3.38

图4

LGDP基因的精确定位 A: LGDP基因的定位; B: LGDP候选基因的转录组测序分析; C: LGDP候选基因的PCR分析和序列比对。N: 天冬酰胺; E: 谷氨酸; A: 丙氨酸; D: 天冬氨酸; M: 蛋氨酸; V: 缬氨酸; H: 组氨酸; TGA: 终止密码子。"

图5

粒形相关基因表达分析 将1B的穗中的转录水平设置为1.0, Bar值表示3个生物学重复的平均值±SD; **表示在0.01概率水平差异显著。"

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