水稻垩白基因<em>Chalk</em>5功能标记的开发与应用

doi:10.7668/hbnxb.2017.01.001

华北农学报 ›› 2017, Vol. 32 ›› Issue (1): 1-8. doi: 10.7668/hbnxb.2017.01.001

所属专题： 水稻；生物技术；

水稻垩白基因Chalk5功能标记的开发与应用

朱金燕^1,2, 王晴晴³, 王军^1,2, 范方军^1,2, 李文奇^1,2, 王芳权^1,2, 王绪鹏⁴, 仲维功^1,2, 杨杰^1,2

1. 江苏省农业科学院粮食作物研究所, 国家水稻改良中心南京分中心, 江苏南京 210014;
2. 扬州大学江苏省粮食作物现代产业技术协同创新中心, 江苏扬州 225009;
3. 苏州农业职业技术学院, 江苏苏州 215008;
4. 扬州大学生物科学与技术学院, 江苏扬州 225009

收稿日期:2016-10-10 出版日期:2017-02-28
通讯作者: 杨杰(1969-),男,四川南充人,研究员,博士,主要从事水稻遗传育种研究。
作者简介:朱金燕(1983-),女,江苏南通人,副研究员,博士,主要从事水稻遗传育种研究。
基金资助:
国家自然科学基金项目（31400175）；江苏省自然科学基金项目（BK20130725）；江苏省现代农业研发项目（BE2015355）；江苏科技成果转化资金（BA2014134）；公益性行业科研专项（201303102）；扬州市农业前瞻性研究项目（YZ2014165）

Development and Application of Two Functional Markers for Grain Chalkiness Gene Chalk5 in Rice

ZHU Jinyan^1,2, WANG Qingqing³, WANG Jun^1,2, FAN Fangjun^1,2, LI Wenqi^1,2, WANG Fangquan^1,2, WANG Xupeng⁴, ZHONG Weigong^1,2, YANG Jie^1,2

1. Institute of Food Crops, Jiangsu Provine Academy of Agricultural Sciences, Nanjing Branch of Chinese National Center for Rice Improvement, Nanjing 210014, China;
2. Jiangsu Co-innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China;
3. Suzhou Polytechnic Institute of Agriculture, Suzhou 215008, China;
4. College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, China

Received:2016-10-10 Published:2017-02-28

摘要/Abstract

摘要： 水稻垩白是衡量水稻外观品质优劣的重要指标之一，降低稻米的垩白是培育优质水稻品种的前提。Chalk5是一个能显著降低稻米垩白从而提高稻米外观品质的主效基因。为提高垩白基因Chalk5在育种中的选择效率，在前人克隆的基础上，根据其启动子区域存在的2个功能突变位点，分别设计等位基因特异PCR（ARMS-PCR）引物，最终筛选出2对Chalk5基因的功能标记Chalk12、NChalk12和Chalk22、NChalk22，通过PCR产物测序的方法对这2对标记的检查结果进行了验证，说明Chalk12、NChalk12和Chalk22、NChalk22可以准确鉴定出Chalk5的不同基因型。利用该标记对不同来源的43份籼稻资源、江苏2007-2013年审定的65份粳稻品种和太湖流域179份粳稻地方资源进行了Chalk5基因型检测，筛选到携带chalk5低垩白率等位基因的籼稻资源20份，太湖流域粳稻地方资源仅8份，其余259份粳型资源中均不携带chalk5低垩白率等位基因，这也说明chalk5主要分布在籼型水稻资源中，在粳型资源中分布很少。

关键词: 水稻, 垩白, Chalk5, 等位基因特异PCR, 功能标记Q78

Abstract: Grain chalkiness is one of the important indicators to evaluate the quality of rice appearance. Reducing grain chalkiness is the basis for high quality rice breeding. Chalk5 is a major quantitative trait locus which has been cloned and can influence grain chalkiness. In order to increase the selection efficiency of Chalk5 in breeding, six pairs of amplification refractory mutation system PCR (ARMS-PCR) primers had been designed basing on two functional mutations in promoter region. Finally, two pairs of functional markers,Chalk12/NChalk12 and Chalk22/NChalk22 had been selected out. The sequenced results of target region were consisted with the PCR results which verified the detection accuracy of the newly designed markers. Chalk12/NChalk12 and Chalk22/NChalk22 were employed to genotype 43 indica rice varieties, 65 japonica varieties certificated in Jiangsu from 2007 to 2013, and 179 local japonica cultivars from Taihu Basin. The results showed that only 20 indica rice varieties and 8 local japonica cultivars from Taihu Basin carried chalk5 gene, while the other 259 rice varieties carried Chalk5 gene. All these indicated that chalk5 mainly existing in indica germplasm.

Key words: Rice, Chalkiness, Chalk5, Amplification refractory mutation system PCR, Functional marker

中图分类号:

朱金燕, 王晴晴, 王军, 范方军, 李文奇, 王芳权, 王绪鹏, 仲维功, 杨杰. 水稻垩白基因Chalk5功能标记的开发与应用[J]. 华北农学报, 2017, 32(1): 1-8. doi: 10.7668/hbnxb.2017.01.001.

ZHU Jinyan, WANG Qingqing, WANG Jun, FAN Fangjun, LI Wenqi, WANG Fangquan, WANG Xupeng, ZHONG Weigong, YANG Jie. Development and Application of Two Functional Markers for Grain Chalkiness Gene Chalk5 in Rice[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(1): 1-8. doi: 10.7668/hbnxb.2017.01.001.

http://www.hbnxb.net/CN/Y2017/V32/I1/1

参考文献

[1] Fitzgerald M, Mccouch S, Hall R. Not just a grain of rice:the quest for quality[J]. Trends in Plant Science, 2009, 14(3):133-139.
[2] Siebenmorgen T, Grigg B, Lanning S. Impacts of preharvest factors during kernel development on rice quality and functionality[J]. Annual Review of Food Science and Technology,2013,4(4):101-115.
[3] Bowles D. Towards increased crop productivity and quality[J]. Current Opinion in Biotechnology, 2012, 23(2):202-203.
[4] Zhang Q. Strategies for developing Green Super Rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(104):16402-16409.
[5] Xing Y, Zhang Q. Genetic and molecular bases of rice yield[J]. Annual Review of Plant Biology, 2010, 61(1):421-442.
[6] Fernie A, Tadmor Y, Zamir D. Natural genetic variation for improving crop quality[J]. Current Opinion in Plant Biology, 2006, 9(2):196-202.
[7] Kang H, Park S, Matsuoka M, et al. White-core endosperm floury endosperm-4 in rice is generated by knockout mutations in the C-type pyruvate orthophosphate dikinase gene (OsPPDKB)[J]. The Plant Journal:for Cell and Molecular Biology, 2005, 42(6):901-911.
[8] Singh N, Kaur L, Sandhu K S, et al. Relationships between physicochemical, morphological, thermal, rheological properties of rice starches[J]. Food Hydrocolloids, 2006, 20(4):532-542.
[9] Fujita N, Yoshida M, Kondo T, et al. Characterization of SSIIIa-deficient mutants of rice:the function of SSIIIa and pleiotropic effects by SSIIIa deficiency in the rice endosperm[J]. Plant Physiology, 2007, 144(4):2009-2023.
[10] Yamakawa H, Hirose T, Kuroda M, et al. Comprehensive expression profiling of rice grain filling-related genes under high temperature using DNA microarray[J]. Plant Physiology, 2007, 144(1):258-277.
[11] Satoh H, Omura T. New endosperm mutations induced by chemical mutagens in rice (Oryza sativa L.)[J].Japanese Tournal of Breeding,1981,31(3):316-326.
[12] Tan Y F, Xing Y Z, Li J X, et al. Genetic bases of appearance quality of rice grains in Shanyou 63, an elite rice hybrid[J]. Theoretical and Applied Genetics, 2000, 101(5/6):823-829.
[13] Li J M, Xiao J H, Grandillo S, et al. QTL detection for rice grain quality traits using an interspecific backcross population derived from cultivated Asian (O.sativa L.) and African (O.glaberrima S.) rice[J]. Genome, 2004, 47(4):697-704.
[14] 顾蕴洁,熊飞,王忠,等.水稻和小麦胚乳发育的比较[J].南京师大学报:自然科学版, 2001, 24(3):65-74.
[15] 李欣,莫惠栋,王安民,等.粳型杂种稻米几个品质性状的遗传表达[J].中国水稻科学, 1999, 13(4):197-204.
[16] 徐富贤,熊洪,朱永川,等.杂交中稻整精米率的影响因子及其间接鉴定方法研究[J].西南农业学报, 2013, 26(1):1-6.
[17] 周少川,李宏,王家生,等.华南籼稻早造稻米蒸煮、外观和碾米品质与食味品质的相关性研究[J].作物学报, 2002, 28(3):397-400.
[18] 刘奇华,蔡建,刘敏,等.两个籼稻品种垩白对稻米蒸煮食味与营养品质的影响[J].中国水稻科学, 2007, 21(3):327-330.
[19] Lanning S B, Siebenmorgen T J, Counce P A, et al. Extreme nighttime air temperatures in 2010 impact rice chalkiness and milling quality[J]. Field Crops Research, 2011, 124(1):132-136.
[20] Fan C, Yu X, Xing Y, et al. The main effects, epistatic effects and environmental interactions of QTLs on the cooking and eating quality of rice in a doubled-haploid line population[J]. Theoretical and Applied Genetics,2005,110(8):1445-1452.
[21] Lin S, Tsai J, Hsiao C, et al. Crystal structure of a membrane-embedded H⁺-translocating pyrophosphatase[J]. Nature, 2012, 484(7394):399-403.
[22] Yamakawa H, Ebitani T, Terao T. Comparison between locations of QTLs for grain chalkiness and genes responsive to high temperature during grain filling on the rice chromosome map[J]. Breeding Science, 2008, 58(3):337-343.
[23] Qin Y, Kim S M, Sohn J K. Genetic analysis and QTL mapping for grain chalkiness characteristics of brown rice (Oryza sativa L.)[J]. Genes & Genomics, 2009, 31(2):155-164.
[24] Zhou L J, Chen L M, Jiang L, et al. Fine mapping of the grain chalkiness QTL qPGWC-7 in rice (Oryza sativa L.)[J]. Theoretical and Applied Genetics, 2009, 118(3):581-590.
[25] Song X J, Huang W, Shi M, et al. A QTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase[J]. Nature Genetics, 2007, 39(5):623-630.
[26] Li Y B, Fan C C, Xing Y Z, et al. Chalk5 encodes a vacuolar H⁺-translocating pyrophosphatase influencing grain chalkiness in rice[J]. Nature Genetics, 2014, 46(4):398.
[27] Ezcurra I, Ellerström M, Wycliffe P, et al. Interaction between composite elements in the napA promoter:both the B-box ABA-responsive complex and the RY/G complex are necessary for seed-specific expression[J]. Plant Molecular Biology, 1999, 40(4):699-709.
[28] Ezcurra I, Wycliffe P, Nehlin L, et al. Transactivation of the Brassica napus napin promoter by ABI3 requires interaction of the conserved B2 and B3 domains of ABI3 with different cis-elements:B2 mediates activation through an ABRE, whereas B3 interacts with an RY/g-box[J].PubMed,2000, 24(1):57-66.
[29] Delgado-Alvarado A, Walker R, Leegood R. Phosphoenolpyruvate carboxykinase in developing pea seeds is associated with tissues involved in solute transport and is nitrogen-responsive[J]. Plant, Cell & Environment, 2007, 30(2):225-235.
[30] Gowik U, Burscheidt J, Akyildiz M, et al. cis-Regulatory elements for mesophyll-specific gene expression in the C4 plant Flaveria trinervia, the promoter of the C4 phosphoenolpyruvate carboxylase gene[J].Plant Cell, 2004, 16(5):1077.
[31] Walker R, Battistelli A, Moscatello S, et al. Metabolism of the seed and endocarp of cherry (Prunus avium L.) during development[J]. Plant Physiology and Biochemistry,2011, 49(8):923-930.
[32] Ye S, Dhillon S, Ke X, et al. An efficient procedure for genotyping single nucleotide polymorphisms[J]. Nucleic Acids Research, 2001, 29(17):8.
[33] Dellaporta S L, Wood J, Hicks J B. A plant DNA minipreparation:Version Ⅱ[J]. Plant Molecular Biology Reporter, 1983, 1(4):19-21.
[34] 万建民.作物分子设计育种[J].作物学报, 2006, 32(3):455-462.
[35] 黎裕,王建康,邱丽娟,等.中国作物分子育种现状与发展前景[J].作物学报, 2010, 36(9):1425-1430.
[36] Fjellstrom R, Conaway-Bormans C A, Mcclung A M, et al. Development of DNA markers suitable for marker assisted selection of three Pi genes conferring resistance to multiple Pyricularia grisea pathotypes[J]. Crop Science, 2004, 44(5):1790-1798.
[37] 李军,李白,高荣村.利用四引物扩增受阻突变体系PCR技术检测水稻光温敏核不育基因 p/tms12-1[J].中国水稻科学, 2014, 28(4):442-446.
[38] 王军,杨杰,朱金燕,等.稻瘟病抗病基因 Pi-kh 功能标记的开发及江苏粳稻品种中Pi-kh的变异[J].中国水稻科学, 2014, 28(2):141-147.
[39] Andersen J R, Lübberstedt T. Functional markers in plants[J]. Trends in Plant Science, 2003, 8(11):554-560.

选择文件类型/文献管理软件名称

选择包含的内容

水稻垩白基因Chalk5功能标记的开发与应用

Development and Application of Two Functional Markers for Grain Chalkiness Gene Chalk5 in Rice

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	李霞, 罗丽卉, 周娅, 杨定清, 王棚, 李森. 秸秆还田对水稻-油菜轮作体系土壤活性有机碳组分及酶活性的影响[J]. 华北农学报, 2022, 37(5): 124-131.
[2]	王亚, 王越涛, 申关望, 王付华, 王生轩, 白涛, 尹海庆. *聚合R基因Pigm和非R基因bsr-d1改良水稻稻瘟病抗性*[J]. 华北农学报, 2022, 37(5): 157-165.
[3]	周丽平, 赵秋, 张新建, 宁晓光, 袁亮, 李燕婷, 赵秉强. 新型增效复合肥料对水稻养分吸收和产量的影响[J]. 华北农学报, 2022, 37(2): 112-120.
[4]	梁玉刚, 周晶, 余政军, 李静怡, 黄璜, 赵正洪. 稻鸭共育对直播水稻田间杂草群落组成及多样性的影响[J]. 华北农学报, 2022, 37(2): 160-170.
[5]	韩政宏, 段宇轩, 徐善斌, 王敬国, 刘化龙, 杨洛淼, 贾琰, 辛威, 郑洪亮, 邹德堂. *利用CRISPR/Cas9技术敲除GS3和GS9基因改良水稻粒型性状*[J]. 华北农学报, 2022, 37(2): 9-17.
[6]	刘俊峰, 李漪濛, 梁超, 周婵婵, 王术, 贾宝艳, 黄元财, 王岩, 王韵. 施氮方式与行距配置对水稻冠层结构及产量的影响[J]. 华北农学报, 2022, 37(1): 77-85.
[7]	静莉丽, 彭廷, 赵亚帆, 赵帅兵, 王童童, 李源, 程远, 杜彦修, 张静, 孙红正, 赵全志. 灌浆充实调节剂对大穗型水稻弱势籽粒灌浆的调控效应[J]. 华北农学报, 2022, 37(1): 86-94.
[8]	徐令旗, 郭晓红, 张佳柠, 赵洋, 李晓蕾, 刘绍峰, 崔致远, 安懿亮, 吕艳东. 不同有机肥对旱直播水稻品质的影响[J]. 华北农学报, 2022, 37(1): 137-146.
[9]	陈子琪, 王伟苹, 赵宏强, 王皓, 韩笑, 杨洛淼, 辛威, 刘化龙, 郑洪亮, 王敬国. 利用籼粳交RIL群体进行水稻发芽期与芽期耐冷性QTL分析[J]. 华北农学报, 2022, 37(1): 50-57.
[10]	董林林, 沈明星, 全坚宇, 闻育琴, 沈园, 陶玥玥, 施林林, 陆长婴. 基肥正位深施对直播水稻产量和化肥利用的影响[J]. 华北农学报, 2021, 36(S1): 222-230.
[11]	张鹏, 吴佩聪, 单颖, 邹刚华, 丁哲利, 钱永德, 赵凤亮. 秸秆还田对热带土壤-水稻种植系统N₂O排放的影响[J]. 华北农学报, 2021, 36(S1): 260-266.
[12]	隆艳喜, 罗雁茹, 竺正航, 廖芷依, 王兰. 四倍体水稻蛋白质含量和谷蛋白基因表达研究[J]. 华北农学报, 2021, 36(6): 78-87.
[13]	魏吉平, 代航, 李振勇, 丁紫苏, 唐凌, 栾鑫, 柯善文, 张向前. *水稻叶早衰突变体osles3的基因定位及其生物信息学分析*[J]. 华北农学报, 2021, 36(6): 45-53.
[14]	彭立功, 龚静, 兰艳, 王锦, 隋晓东, 丁春邦, 李天. 水分胁迫对宜香优2115籽粒淀粉合成及产量的影响[J]. 华北农学报, 2021, 36(5): 77-86.
[15]	李静怡, 王忍, 孟祥杰, 梁玉刚, 龚向胜, 黄璜, 魏海林. 免耕半固态直播对水稻剑叶生理特性的影响[J]. 华北农学报, 2021, 36(5): 87-98.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

水稻垩白基因Chalk5功能标记的开发与应用

Development and Application of Two Functional Markers for Grain Chalkiness Gene Chalk5 in Rice

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价