中国水稻科学 ›› 2018, Vol. 32 ›› Issue (4): 349-356.DOI: 10.16819/j.1001-7216.2018.7131
齐岳翰1,2, 李瑞莉2,3, 王芳2, 刘洪家2,*(), 易可可4, 朱诚1,*()
收稿日期:
2017-11-01
修回日期:
2018-02-06
出版日期:
2018-07-10
发布日期:
2018-07-10
通讯作者:
刘洪家,朱诚
基金资助:
Yuehan QI1,2, Ruili LI2,3, Fang WANG2, Hongjia LIU2,*(), Keke YI4, Cheng ZHU1,*()
Received:
2017-11-01
Revised:
2018-02-06
Online:
2018-07-10
Published:
2018-07-10
Contact:
Hongjia LIU, Cheng ZHU
摘要: 【目的】本研究旨在鉴定和克隆水稻温敏转绿新基因,揭示其参与叶绿体发生发育和光合作用的分子机制,为高光效育种提供理论支撑。【方法】利用辐射诱变的方法,从粳稻品种日本晴中筛选获得叶片黄化突变体osv15,并对其表型、农艺性状和遗传方式进行详细分析。构建了突变体与Kasalath的F2群体,利用多态性分子标记对目的基因进行定位和测序分析。【结果】osv15幼苗期在22℃低温下叶片黄化,叶绿素含量仅为野生型的10%,光化学效率下降,叶绿体结构异常;随着温度的升高,osv15的叶色由黄转绿,30℃时叶绿素含量恢复到野生型的68%,光化学效率和叶绿体发育与野生型相近。在自然环境下,osv15突变体从苗期至成熟期均表现为叶片黄化,且株高、分蘖数和产量等农艺性状与野生型相比差异显著。遗传分析表明osv15突变体的表型由一对隐性核基因控制。将OsV15基因定位到第6染色体多态性标记S4和S5之间84 kb的区间内,定位区间测序发现突变体中编码分子伴侣蛋白的基因Cpn60β1(LOC_Os06g02380)发生单碱基缺失。【结论】osv15是一个新的水稻温敏转绿突变体,Cpn60β1可能为突变基因。
中图分类号:
齐岳翰, 李瑞莉, 王芳, 刘洪家, 易可可, 朱诚. 水稻温敏转绿突变体osv15的鉴定和遗传分析[J]. 中国水稻科学, 2018, 32(4): 349-356.
Yuehan QI, Ruili LI, Fang WANG, Hongjia LIU, Keke YI, Cheng ZHU. Identification and Genetic Analysis of Thermo-sensitive Mutant osv15 in Rice[J]. Chinese Journal OF Rice Science, 2018, 32(4): 349-356.
标记名称 Marker name | 正向引物(5′-3′) Forward primer(5′-3′) | 反向引物(5′-3′) Reverse primer(5′-3′) | 所在BAC BAC location |
---|---|---|---|
S1 | TAACACGCACTCAAAACGTG | TGCCGACTCTCATTGATCTG | AP001552 |
S2 | CAACCCCTAACGACCATAGC | CACATGCATGACACGATCAA | AP001552 |
S3 | CCAACTCTACATGCTCCTTCTG | CCCCAGAGAAGATCTCCTAGC | AP001389 |
S4 | TCCTCACCTCCTCCTCCTTT | CACAAGCTTCTCTCTTCCCTTC | AP001389 |
S5 | AGAGGCAATGTCGCAGTATG | GGTGGTTCTGCTCCCATTTA | AP002837 |
S6 | ACAACTTCGCTTCTCGGTTG | GTACCTGACAGAGGCGATCC | AP002838 |
表1 用于精细定位的多态性STS标记
Table 1 Polymorphic STS markers used in fine mapping.
标记名称 Marker name | 正向引物(5′-3′) Forward primer(5′-3′) | 反向引物(5′-3′) Reverse primer(5′-3′) | 所在BAC BAC location |
---|---|---|---|
S1 | TAACACGCACTCAAAACGTG | TGCCGACTCTCATTGATCTG | AP001552 |
S2 | CAACCCCTAACGACCATAGC | CACATGCATGACACGATCAA | AP001552 |
S3 | CCAACTCTACATGCTCCTTCTG | CCCCAGAGAAGATCTCCTAGC | AP001389 |
S4 | TCCTCACCTCCTCCTCCTTT | CACAAGCTTCTCTCTTCCCTTC | AP001389 |
S5 | AGAGGCAATGTCGCAGTATG | GGTGGTTCTGCTCCCATTTA | AP002837 |
S6 | ACAACTTCGCTTCTCGGTTG | GTACCTGACAGAGGCGATCC | AP002838 |
图1 不同时期野生型(WT)和突变体表型 A–22℃培养箱生长14 d;B–30℃培养箱生长9 d;C–成熟期(11月);D–成熟期(7月)。标尺=3 cm。
Fig. 1. Phenotype of the wild type (WT) and mutant at different growth stages. A, 14 d-old seedlings at 22℃; B, 9 d-old seedlings at 30℃; C, At maturity stage in November; D, At maturity stage in July. Bar=3 cm.
材料 Material | 株高 Plant height / cm | 分蘖数 Tiller number | 有效穗数 No. of productive panicles | 穗长 Panicle length / cm | 叶长 Leaf length / cm | 叶宽 Leaf width / cm | 结实率 Seed setting rate / % | 千粒重 1000-grain weight / g |
---|---|---|---|---|---|---|---|---|
WT | 87.3±2.0 | 15.8±2.2 | 14.3±2.1 | 21.5±0.6 | 31.5±1.3 | 1.3±0.1 | 91.4±2.1 | 25.8±0.6 |
osv15 | 80.2±1.9** | 8.6±1.4** | 6.8±1.0** | 20.1±1.0 | 29.9±1.5 | 1.2±0.1 | 80.1±4.0** | 22.8±1.8** |
表2 野生型和突变体农艺性状分析
Table 2 Agronomic traits of the wild type and mutant.
材料 Material | 株高 Plant height / cm | 分蘖数 Tiller number | 有效穗数 No. of productive panicles | 穗长 Panicle length / cm | 叶长 Leaf length / cm | 叶宽 Leaf width / cm | 结实率 Seed setting rate / % | 千粒重 1000-grain weight / g |
---|---|---|---|---|---|---|---|---|
WT | 87.3±2.0 | 15.8±2.2 | 14.3±2.1 | 21.5±0.6 | 31.5±1.3 | 1.3±0.1 | 91.4±2.1 | 25.8±0.6 |
osv15 | 80.2±1.9** | 8.6±1.4** | 6.8±1.0** | 20.1±1.0 | 29.9±1.5 | 1.2±0.1 | 80.1±4.0** | 22.8±1.8** |
图2 不同温度下野生型和突变体光合色素含量 *表示野生型与突变体在0.05水平上差异显著。
Fig. 2. Photosynthetic pigment contents in leaves of the wild type(WT) and mutant at different temperatures. *Difference between WT and mutant was significant at 0.05 level.
图3 野生型和突变体叶绿体超微结构 A–22℃条件下野生型;B–22℃条件下突变体;C–30℃条件下野生型;D–30℃条件下突变体。G–基粒;O–嗜锇粒。
Fig. 3. Chloroplast ultrastructure of the wild type and mutant. A, The wild type under 22℃; B, Mutant under 22℃; C, The wild type under 30℃; D, Mutant under 30℃. G, Granum; O, Osmiophilic globule.
图4 不同温度下野生型和突变体叶绿素荧光动力学参数 A–最大光化学效率(Fv/Fm);B–实际光化学效率(ΦPSII);C–光化学猝灭系数(qP)。
Fig. 4. Chlorophyll fluorescence kinetics parameters of the wild type(WT) and mutant at different temperatures. A, Maximum photochemical efficiency(Fv/Fm); B, Actual photochemical efficiency(ΦPSII); C, Photochemical quenching coefficient(qP).
组合 Combination | 绿苗株数 Number of green-leaf plants | 黄苗株数 Number of yellow leaf plants | 总株数 Total number of plants | 分离比 Segregation | χ2 |
---|---|---|---|---|---|
osv15/Nipponbare osv15/Kasalath | 310 279 | 95 88 | 405 367 | 3∶1 3∶1 | 0.53 0.29 |
表3 突变体后代叶色性状的分离分析
Table 3 Segregation of leaf color of progenies derived from mutant.
组合 Combination | 绿苗株数 Number of green-leaf plants | 黄苗株数 Number of yellow leaf plants | 总株数 Total number of plants | 分离比 Segregation | χ2 |
---|---|---|---|---|---|
osv15/Nipponbare osv15/Kasalath | 310 279 | 95 88 | 405 367 | 3∶1 3∶1 | 0.53 0.29 |
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