水稻黄绿叶突变体djyg的遗传分析与基因定位

doi:10.3969/j.issn.1001G7216.2015.06.006

中国水稻科学 ›› 2015, Vol. 29 ›› Issue (6): 601-609.DOI: 10.3969/j.issn.1001G7216.2015.06.006

水稻黄绿叶突变体djyg的遗传分析与基因定位

李智强^1,^2,³, 朱丹^1,^2,³, 王志龙^1,², 丁波³, 王国梁^1,^2,^3,^*()

¹湖南农业大学农学院,长沙 410128
²作物基因工程湖南省重点实验室,长沙 410128
³中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室, 北京100193

收稿日期:2015-06-18 修回日期:2015-07-09 出版日期:2015-10-25 发布日期:2015-11-10
通讯作者: 王国梁
作者简介:
^*通讯录作者：E-mail:wang.620@osu.edu
基金资助:
基金项目: 国家自然科学基金资助项目(31371928)教育部 “创新团队发展计划”资助项目(IRT1239)湖南省高校科技创新团队项目(2012)

Genetic Analysis and Gene Mapping of a Yellow-Green Leaf Mutant djyg in Rice

Zhi-qiang LI^1,^2,³, Dan ZHU^1,^2,³, Zhi-long WANG^1,², Bo DING³, Guo-liang WANG^1,^2,^3,^*

¹ College of Agronomy, Hunan Agricultural University, Changsha 410128, China
Crop Gene Engineering Key Laboratory of Hunan Province, Changsha 410128, China
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2015-06-18 Revised:2015-07-09 Online:2015-10-25 Published:2015-11-10
Contact: Guo-liang WANG
About author:
^*Corresponding author：E-mail:wang.620@osu.edu

摘要/Abstract

摘要：

在粳稻品种Dongjin大田种植过程中,发现一个黄绿叶自然突变体,命名为djyg。该突变体在苗期表现明显的黄绿叶表型,抽穗以后,叶色逐渐恢复正常。叶绿素含量测定结果表明,在苗期、分蘖盛期及抽穗期叶绿素b的含量分别下降53%、62%、36%。电镜结果表明,分蘖期突变体中基粒、类囊体垛堆凌乱、排列疏松,类囊体基质较为稀薄。qRT-PCR结果证实,PORA、Cab1R、PsbA的表达量在突变体中均较野生型明显下调。遗传分析结果表明,黄绿叶突变体djyg由一对隐性主效核基因控制,图位克隆确定该候选基因为编码叶绿素合成酶基因YGL1的一个新等位基因。该突变体未影响植株的主要农艺性状,可作为一个理想的表型标记应用于杂交稻育种工作中。

关键词: 水稻, 黄绿叶, 突变体, 图位克隆

Abstract:

A spontaneous yellow-green mutant was obtained from the japonica rice cultivar Dongjin in the field, designated as djyg. The mutant showed obvious yellow-green leaf phenotype at the seedling stage but the chlorotic leaves returned to green after the heading stage. Consistent with the phenotype, the chlorophyll b content of the mutant was reduced by 53%, 62% and 36% at the seedling, tillering and heading stages, respectively, compared with that of the wild type (WT). At the tillering stage, we observed that the granas and thylakoid stacks displayed a mess and loose structure, as well as less density in the mutant djyg compared with the WT under a transmission electron microscopy. The transcriptional levels of PORA, Cab1R, PsbA were also decreased in the mutant djyg. Genetic analysis and map-based cloning demonstrated that the phenotype of the mutant was controlled by a major recessive nuclear gene, which is an allele of chlorophyll synthase YGL1. The mutant djyg can be used as a phenotypic marker in rice breeding program since the main agronomic traits of the mutant djyg are normal compared with those of the WT.

Key words: Oryza sativa L., yellow-green leaf, mutant, map-based cloning

李智强, 朱丹, 王志龙, 丁波, 王国梁. 水稻黄绿叶突变体djyg的遗传分析与基因定位[J]. 中国水稻科学, 2015, 29(6): 601-609.

Zhi-qiang LI, Dan ZHU, Zhi-long WANG, Bo DING, Guo-liang WANG. Genetic Analysis and Gene Mapping of a Yellow-Green Leaf Mutant djyg in Rice[J]. Chinese Journal OF Rice Science, 2015, 29(6): 601-609.

图/表 9

参考文献 28

[1]	Khush G S.Feeding five billion rice consumers-The role of rice breeding.Adv Rice Sci, 2004: 3-15.
[2]	Wang P R, Gao J X, Wan C M, et al.Divinyl chlorophyll (ide) a can be converted to monovinyl chlorophyll (ide) a by a divinyl reductase in rice.Plant Physiol, 2010, 153: 994-1003.
[3]	Kusumi K, Sakata C, Nakamura T, et al.A plastid protein NUS1 is essential for build-up of the genetic system for early chloroplast development under cold stress conditions.Plant J, 2011, 68(6): 1039-1050.
[4]	Su N, Hu M L, Wu D X, et al.Disruption of a rice pentatricopeptide repeat protein causes a seedling-specific albino phenotype and its utilization to enhance seed purity in hybrid rice production.Plant Physiol, 2012, 59(1): 227-238.
[5]	Shu Q Y, Liu G F, Xia Y W.Temperature-sensitve leaf color mutation in rice (Oryza sativa L.).Acta Agric Nucl Sini, 1995, 10: 6-10.
[6]	邓晓娟,张海清,王悦,等. 水稻叶色突变基因研究进展. 杂交水稻, 2012,27(5): 9-14.
[7]	Deng X J, Zhang H Q, Wang Y, et al.Mapped clone and functional analysis of leaf-color gene ygl7 in a rice hybrid (Oryza sativa L. ssp. indica).PloS One, 2014, 9(6): e99564.
[8]	Zhou Y, Gong Z Y, Yang Z F, et al.Mutation of the light induced yellow leaf 1 gene, which encodes a geranyl reductase, affects chlorophyll biosynthesis and light sensitivity in rice.PloS One, 2013(8): e75299.
[9]	Tian X Q, Ling Y H, Fang L K, et al.Gene cloning and functional analysis of yellow green leaf 3(ygl3) gene during the whole-plant growth stage in rice.Genes Genom, 2013, 35: 87-93.
[10]	Jiang H W, Li M R, Liang N T, et al.Molecular cloning and function analysis of the stay green gene in rice.Plant J, 2007, 52: 197-209.
[11]	董青,张迎信,张振华,等.水稻黄绿叶突变体W390的遗传分析和基因定位.中国水稻科学, 2015, 29(3): 241-249.
[12]	Huang X Q, Wang P R, Zhao H X, et al.Genetic analysis and molecular mapping of a novel chlorophyll-deficit mutant gene in rice.Rice Sci, 2008, 15: 7-12.
[13]	Wang P R, Ma X Z, Li C M, et al.Comparative analysis of three mutants of divinyl reductase gene in rice.Sci Agric Sin(Chinese Version), 2013, 46: 1305-1313.
[14]	Sakuraba Y, Rahman M L, Cho S H, et al.The rice faded green leaf locus encodes protochlorophyllide oxidoreductase B and is essential for chlorophyll synthesis under high light conditions.Plant J, 2013, 74: 122-133.
[15]	Wang P R, Wang B, Sun X Q, et al.Fine mapping and physiological characteristics of a green-revertible albino gene gra75 in rice.Sci Agric Sin (Chinese Version), 2013, 46: 225-232.
[16]	Wu Z, Zhang X, He B, et al.A chlorophyll-deficient rice mutant with impaired chlorophyllide esterification in chlorophyll biosynthesis.Plant Physiol, 2007, 145(1): 29-40.
[17]	Wang P, Li C, Wang Y, et al.Identification of a geranylgeranyl reductase gene for chlorophyll synthesis in rice.Springer Plus, 2014, 3(1): 201.
[18]	Wan C, Li C, Ma X, et al.GRY79 encoding a putative metallo-β-lactamase-trihelix chimera is involved in chloroplast development at early seedling stage of rice.Plant Cell Rep, 2015: 1-11.
[19]	Harmut K.Chlorophyls and carotenoids: Pigments of photosynthetic biomembranes.Methods Enzymol, 1987, 148(34): 350-382.
[20]	Nie H, Zhao C, Wu G, et al.SR1, a calmodulin-binding transcription factor, modulates plant defense and ethylene-induced senescence by directly regulating NDR1 and EIN3.Plant Physiol, 2012, 158: 1847-1859.
[21]	Pogers S O, Bendich A J.Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues.Plant Mol Biol, 1985, 5: 69-76.
[22]	Zhang H T, Li J J, Yoo J H, et al.Rice Chlorina-1 and Chlorina-9 encode ChlD and ChlI subunits of Mg-chelatase, a key enzyme for chlorophyll synthesis and chloroplast development.Plant Mol Biol, 2006, 62: 325-337.
[23]	Sun X Q, Wang B, Xiao Y H, et al.Genetic analysis and fine mapping of gene ygl98 for yellow-green leaf of rice.Acta Agrono Sinica(Chinese Version), 2011, 37: 991-997.
[24]	Rüdiger W.Biosynthesis of chlorophyll b and the chlorophyll cycle.Phot Res, 2002, 74:187-193.
[25]	Stenbaek A, Jensen P E.Redox regulation of chlorophyll biosynthesis.Phyto Chem, 2010, 71: 853-859.
[26]	Tang Y L, Huang J F, Wang R C.Change law of hyperspectral data in related with chlorophyll and carotenoid in rice at different developmental stages.Rice Sci, 2004, 11: 274-282.
[27]	Yasuhito S, Md L R, Cho S H, et al.The rice faded green leaf locus encodes protochlorophyllide oxidoreductase B and is essential for chlorophyll synthesis under high light conditions.Plant J, 2013, 74: 122-133.
[28]	Baumgartner B J, Rapp J C, Mullet J E, et al.Plastid genes encoding the transcription/translation apparatus are differentially transcribed early in barley (hordeum vulagare) chloroplast development (evidence for selective stabilization of psbA mRNA).Plant Physiol, 1993, 101: 781-791.

引物名称 Primer name	正向引物(5'-3') Forward primer (5'-3')	反向引物(5'-3') Reverse primer (5'-3')
HEMA1	CGCTATTTCTGATGCTATGGGT	GAGCACAGCAAAATCCTAGACG
PORA	TGTACTGGAGCTGGAACAACAA	GAGCACAGCAAAATCCTAGACG
V1(NUS1)	TGGAGGTCGGGACAGAGGA	CGAGGAGCACCACCATCAC
Cab1R	AGATGGGTTTAGTGCGACGAG	TTTGGGATCGAGGGAGTATTT
CAO1	GATCCATACCCGATCGACAT	CGAGAGACATCCGGTAGAGC
RbcL	CTTGGCAGCATTCCGAGTAA	ACAACGGGCTCGATGTGATA
OsPPR1	CTAAGACCGAATGACAAATGC	GCACTGCCAACAAGAATACC
V2	CGACAAGCAGAGCGAAGCG	AGGTTGCTGCTCCTTGAATGT
Cab2R	TGTTCTCCATGTTCGGCTTCT	GCTACGGTCCCCACTTCACT
PsaA	GCGAGCAAATAAAACACCTTTC	GTACCAGCTTAACGTGGGGAG
PsbA	CCCTCATTAGCAGATTCGTTTT	ATGATTGTATTCCAGGCAGAGC
SPP	CGGAGAGGAAACATAATGAC	ATAGGCATTTGTCTTTGTCTC

引物名称 Primer name	正向引物(5'-3') Forward primer (5'-3')	反向引物(5'-3') Reverse primer (5'-3')
HEMA1	CGCTATTTCTGATGCTATGGGT	GAGCACAGCAAAATCCTAGACG
PORA	TGTACTGGAGCTGGAACAACAA	GAGCACAGCAAAATCCTAGACG
V1(NUS1)	TGGAGGTCGGGACAGAGGA	CGAGGAGCACCACCATCAC
Cab1R	AGATGGGTTTAGTGCGACGAG	TTTGGGATCGAGGGAGTATTT
CAO1	GATCCATACCCGATCGACAT	CGAGAGACATCCGGTAGAGC
RbcL	CTTGGCAGCATTCCGAGTAA	ACAACGGGCTCGATGTGATA
OsPPR1	CTAAGACCGAATGACAAATGC	GCACTGCCAACAAGAATACC
V2	CGACAAGCAGAGCGAAGCG	AGGTTGCTGCTCCTTGAATGT
Cab2R	TGTTCTCCATGTTCGGCTTCT	GCTACGGTCCCCACTTCACT
PsaA	GCGAGCAAATAAAACACCTTTC	GTACCAGCTTAACGTGGGGAG
PsbA	CCCTCATTAGCAGATTCGTTTT	ATGATTGTATTCCAGGCAGAGC
SPP	CGGAGAGGAAACATAATGAC	ATAGGCATTTGTCTTTGTCTC

引物名称 Primer name		正向引物(5'-3') Forward primer (5'-3')	反向引物(5'-3') Reverse primer (5'-3')	产物大小(bp) Product size(bp)
HM-1	InDel	TTAGGCACACTTGGCAGACA	AGAGATGACACGGTTTGGGG	200
HM-2	InDel	GGCATAAGAGGTACTTCTGGGA	TGAAGGCACAATTTAGGTGAACG	183
HM-3	InDel	CCCTGCAATGCAAACGAGAG	TCCTCATTCGATTAGTCGGCG	168
HM-4	InDel	AATTGGGCCATCTGTACGCA	ATTTTTGCCCTTCAACCGCC	176
HM-5	InDel	CCTGGGTTAAGGCAGGTTCG	AGAGTGAGTACCACCACCGTA	204
HM-6	InDel	GGGGTCGCGTGGAAAATTAC	TCCTACCGTGTGGTTACGTG	249
HM-7	InDel	AGTACGCTTCGTGTGGATGG	GGAAGTGCCAGCTCATGATT	237
HM-8	InDel	TCGACATATACAGCGCGGAC	TAGGGAGTACGCTCCCATGT	235
HM-9	InDel	TGCCTATGCTTCCTTTCGTGA	CACATGTGCAAACGTGTGAT	165
HM-10	InDel	TCCCGCCATGGTTATGCTTT	TACCGAGTGCAGGGTAGTCA	225
HM-11	InDel	CATCCGGTCAAACGTCCGAT	GCCGGCCTGCTATGTTTTTA	115
HM-12	InDel	TCCATTTCGGCTCTAGTTGTG	TATAAGGGCTATGGCTGGTTC	123
HM-13	InDel	GCCTCGTACCATCGGATACTCT	ATTTCCATACGCGGTTGCTC	112
HM-14	InDel	ACTACGAATCTGGACATACATA	AGAGTGAGAAGCACATATTGTA	156
HM-15	InDel	CTTCGTGCTTACGGTGGCG	GGCATGACAGGCAAACTGC	178
HM-16	InDel	CCTACCCTCCAACCCATAC	ACAAAGCCGACGCTAATCT	168
HM-17	InDel	AGGGTCGTCGGAACAAGG	TGCCGTAGGGCGTCATC	250
HM-18	InDel	GCTGTCCTTCTTATAGGCG	CGGAGGCATAGCTTGC	177
HM-19	InDel	ACTACTCCGTATGTCCGTGCTA	GTCTACGCATGGGCTTCC	167
HM-20	InDel	TTGATTCAGGTGGTGATGT	CTTGGATTATTTGCGAGAC	200
HM-21	InDel	TACTCCCTCCATTTCAGCATA	GTCATTAACTAGCACCTCTTTT	183
HM-22	InDel	TACTGTTGCACATAGGGTTC	TACTTCAAATGTATGTCCGT	153
HMS1	djyg测序 djyg sequencing	GGGTCAAAAACATCGCGTTA	AACCTAGCTCCACCAACAGT	1309

引物名称 Primer name		正向引物(5'-3') Forward primer (5'-3')	反向引物(5'-3') Reverse primer (5'-3')	产物大小(bp) Product size(bp)
HM-1	InDel	TTAGGCACACTTGGCAGACA	AGAGATGACACGGTTTGGGG	200
HM-2	InDel	GGCATAAGAGGTACTTCTGGGA	TGAAGGCACAATTTAGGTGAACG	183
HM-3	InDel	CCCTGCAATGCAAACGAGAG	TCCTCATTCGATTAGTCGGCG	168
HM-4	InDel	AATTGGGCCATCTGTACGCA	ATTTTTGCCCTTCAACCGCC	176
HM-5	InDel	CCTGGGTTAAGGCAGGTTCG	AGAGTGAGTACCACCACCGTA	204
HM-6	InDel	GGGGTCGCGTGGAAAATTAC	TCCTACCGTGTGGTTACGTG	249
HM-7	InDel	AGTACGCTTCGTGTGGATGG	GGAAGTGCCAGCTCATGATT	237
HM-8	InDel	TCGACATATACAGCGCGGAC	TAGGGAGTACGCTCCCATGT	235
HM-9	InDel	TGCCTATGCTTCCTTTCGTGA	CACATGTGCAAACGTGTGAT	165
HM-10	InDel	TCCCGCCATGGTTATGCTTT	TACCGAGTGCAGGGTAGTCA	225
HM-11	InDel	CATCCGGTCAAACGTCCGAT	GCCGGCCTGCTATGTTTTTA	115
HM-12	InDel	TCCATTTCGGCTCTAGTTGTG	TATAAGGGCTATGGCTGGTTC	123
HM-13	InDel	GCCTCGTACCATCGGATACTCT	ATTTCCATACGCGGTTGCTC	112
HM-14	InDel	ACTACGAATCTGGACATACATA	AGAGTGAGAAGCACATATTGTA	156
HM-15	InDel	CTTCGTGCTTACGGTGGCG	GGCATGACAGGCAAACTGC	178
HM-16	InDel	CCTACCCTCCAACCCATAC	ACAAAGCCGACGCTAATCT	168
HM-17	InDel	AGGGTCGTCGGAACAAGG	TGCCGTAGGGCGTCATC	250
HM-18	InDel	GCTGTCCTTCTTATAGGCG	CGGAGGCATAGCTTGC	177
HM-19	InDel	ACTACTCCGTATGTCCGTGCTA	GTCTACGCATGGGCTTCC	167
HM-20	InDel	TTGATTCAGGTGGTGATGT	CTTGGATTATTTGCGAGAC	200
HM-21	InDel	TACTCCCTCCATTTCAGCATA	GTCATTAACTAGCACCTCTTTT	183
HM-22	InDel	TACTGTTGCACATAGGGTTC	TACTTCAAATGTATGTCCGT	153
HMS1	djyg测序 djyg sequencing	GGGTCAAAAACATCGCGTTA	AACCTAGCTCCACCAACAGT	1309

材料 Material	株高 Plant height /cm	穗长 Panicle length /cm	单株有效穗数 No. of panicles per plant	每穗总粒数 No. of spikelets per panicle	结实率 Seed-setting rate/%	千粒重 1000-grain weight/g
Dongjin	90.68±0.54	18.9±0.5	34±1	118±1	99.24±0.22	22.84±0.71
djyg	90.55±0.45	18.1±0.9	33±1	116±1	98.82±0.21	22.38±0.28

水稻黄绿叶突变体djyg的遗传分析与基因定位

Genetic Analysis and Gene Mapping of a Yellow-Green Leaf Mutant djyg in Rice

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 28

相关文章 15

编辑推荐

Metrics

组合 Cross	总株数 Total number of plants	正常植株 No. of normal plants	黄绿植株 No. of yellow green plants	χ²(3∶1)
9311×djyg	781	592	189	0.23	3.84
Dongjin×djyg	822	629	193	0.93	3.84

[1]	郭展, 张运波. 水稻对干旱胁迫的生理生化响应及分子调控研究进展[J]. 中国水稻科学, 2024, 38(4): 335-349.
[2]	韦还和, 马唯一, 左博源, 汪璐璐, 朱旺, 耿孝宇, 张翔, 孟天瑶, 陈英龙, 高平磊, 许轲, 霍中洋, 戴其根. 盐、干旱及其复合胁迫对水稻产量和品质形成影响的研究进展[J]. 中国水稻科学, 2024, 38(4): 350-363.
[3]	许丹洁, 林巧霞, 李正康, 庄小倩, 凌宇, 赖美玲, 陈晓婷, 鲁国东. OsOPR10正调控水稻对稻瘟病和白叶枯病的抗性[J]. 中国水稻科学, 2024, 38(4): 364-374.
[4]	候小琴, 王莹, 余贝, 符卫蒙, 奉保华, 沈煜潮, 谢杭军, 王焕然, 许用强, 武志海, 王建军, 陶龙兴, 符冠富. 黄腐酸钾提高水稻秧苗耐盐性的作用途径分析[J]. 中国水稻科学, 2024, 38(4): 409-421.
[5]	胡继杰, 胡志华, 张均华, 曹小闯, 金千瑜, 章志远, 朱练峰. 根际饱和溶解氧对水稻分蘖期光合及生长特性的影响[J]. 中国水稻科学, 2024, 38(4): 437-446.
[6]	刘福祥, 甄浩洋, 彭焕, 郑刘春, 彭德良, 文艳华. 广东省水稻孢囊线虫病调查与鉴定[J]. 中国水稻科学, 2024, 38(4): 456-461.
[7]	陈浩田, 秦缘, 钟笑涵, 林晨语, 秦竞航, 杨建昌, 张伟杨. 水稻根系和土壤性状与稻田甲烷排放关系的研究进展[J]. 中国水稻科学, 2024, 38(3): 233-245.
[8]	缪军, 冉金晖, 徐梦彬, 卜柳冰, 王平, 梁国华, 周勇. 过量表达异三聚体G蛋白γ亚基基因RGG2提高水稻抗旱性[J]. 中国水稻科学, 2024, 38(3): 246-255.
[9]	尹潇潇, 张芷菡, 颜绣莲, 廖蓉, 杨思葭, 郭岱铭, 樊晶, 赵志学, 王文明. 多个稻曲病菌效应因子的信号肽验证和表达分析[J]. 中国水稻科学, 2024, 38(3): 256-265.
[10]	朱裕敬, 桂金鑫, 龚成云, 罗新阳, 石居斌, 张海清, 贺记外. 全基因组关联分析定位水稻分蘖角度QTL[J]. 中国水稻科学, 2024, 38(3): 266-276.
[11]	魏倩倩, 汪玉磊, 孔海民, 徐青山, 颜玉莲, 潘林, 迟春欣, 孔亚丽, 田文昊, 朱练峰, 曹小闯, 张均华, 朱春权. 信号分子硫化氢参与硫肥缓解铝对水稻生长抑制作用的机制[J]. 中国水稻科学, 2024, 38(3): 290-302.
[12]	周甜, 吴少华, 康建宏, 吴宏亮, 杨生龙, 王星强, 李昱, 黄玉峰. 不同种植模式对水稻籽粒淀粉含量及淀粉关键酶活性的影响[J]. 中国水稻科学, 2024, 38(3): 303-315.
[13]	关雅琪, 鄂志国, 王磊, 申红芳. 影响中国水稻生产环节外包发展因素的实证研究：基于群体效应视角[J]. 中国水稻科学, 2024, 38(3): 324-334.
[14]	许用强, 姜宁, 奉保华, 肖晶晶, 陶龙兴, 符冠富. 水稻开花期高温热害响应机理及其调控技术研究进展[J]. 中国水稻科学, 2024, 38(2): 111-126.
[15]	吕海涛, 李建忠, 鲁艳辉, 徐红星, 郑许松, 吕仲贤. 稻田福寿螺的发生、危害及其防控技术研究进展[J]. 中国水稻科学, 2024, 38(2): 127-139.