无选择标记基因的优质香稻新品系培育

doi:10.7668/hbnxb.2015.05.015

华北农学报 ›› 2015, Vol. 30 ›› Issue (5): 92-96. doi: 10.7668/hbnxb.2015.05.015

所属专题： 水稻；生物技术；

无选择标记基因的优质香稻新品系培育

冯梦诗¹, 叶俊¹, 侯莉莉¹, 李明¹, 唐丁², 张建祥², 王为云², 徐志豪², 李亚飞², 顾铭洪¹, 程祝宽², 于恒秀¹

1. 扬州大学, 江苏省植物功能基因组学重点实验室, 江苏扬州 225001;
2. 中国科学院遗传与发育生物学研究所, 植物基因组学国家重点实验室, 北京 100101

收稿日期:2015-07-16 出版日期:2015-10-28
通讯作者: 于恒秀(1965-),女,江苏连云港人,博士,教授,主要从事作物遗传育种研究。
作者简介:冯梦诗(1991-),女,江苏扬州人,在读硕士,主要从事作物遗传育种研究。
基金资助:
国家转基因生物新品种培育重大专项(2008ZX08001-006)

Breeding of Marker-free Fragrant Rice Via Transgenic Approach

FENG Meng-shi¹, YE Jun¹, HOU Li-li¹, LI Ming¹, TANG Ding², ZHANG Jian-xiang², WANG Wei-yun², XU Zhi-hao², LI Ya-fei², GU Ming-hong¹, CHENG Zhu-kuan², YU Heng-xiu¹

1. State Key Laboratory of Plant Functional Genomics of the Ministry of Education, Yangzhou University, Yangzhou 225001, China;
2. State Key Laboratory of Plant Genomics and Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China

Received:2015-07-16 Published:2015-10-28

摘要/Abstract

摘要： 香味是稻米食味品质的主要性状之一,为培育无抗性选择标记基因的优质香稻新品系,以携带有超双元载体 O_SBADH2 -RNAi的农杆菌菌株EHA105介导,将 O_SBADH2 基因的RNA干扰结构和潮霉素抗性选择标记(HPT)基因同时导入优质粳稻武育粳3号成熟胚来源的愈伤组织中,经过在含有潮霉素的培养基上筛选和分化,获得了54个独立转化子,其中36个转化子同时含目的基因和 HPT 基因,共转化频率为66.7%;从这些转化子的后代中共筛选获得25个去除 HPT 基因的转化子,去标记频率为69.4%;通过KOH法等4种方法进行香味的鉴定,获得15个具有香味的株系。实时定量PCR分析结果表明,具香味株系中的 O_SBADH2 基因的表达受到了明显抑制,进一步证明了香味的产生是由于 O_SBADH2 基因被干扰而引起。农艺性状调查表明,抑制 O_SBADH2 基因的表达对水稻产量相关性状,包括株高、千粒质量、穗粒数和结实率影响不明显。

关键词: 水稻, 香味, O_SBADH2基因, RNAi, 无选择标记

Abstract: In order to generate marker-free fragrant transgenic rice, O_SBADH2-RNAi and hygromycin phosphotransferase gene(HPT)were transferred to calli derived from a japonica rice cultivar,Wuyujing 3,mediated with a Agrobacterium strain EHA105 harboring a single twin T-DNA binary vector.After selected and regenerated on the media with hygromycin,54 independent transgenic lines were generated.According to PCR results,we got 36 co-transformation lines harboring both HPT gene and O_SBADH2 gene RNAi structure.The frequency of co-transformation was 66.7%.After two generation selections,we got 25 HPT gene free T₂ lines harboring O_SBADH2 gene RNAi structures.The frequency was 69.4%.15 fragrance lines were screened from these transgenic lines using four kind methods detecting fragrance.Real-time quantitative PCR results showed that O_SBADH2 gene expression level was reduced significantly in these transgenic fragrance lines.We found that there was no obvious difference between the transgenic lines and the wild type,in the main agronomy characteristics,including plant height,1000-grain weight,grain number of per spike,and seed setting rate.

Key words: Rice, Fragrant, O_SBADH2, RNAi, Marker-free

中图分类号:

Q78
S511.03

冯梦诗, 叶俊, 侯莉莉, 李明, 唐丁, 张建祥, 王为云, 徐志豪, 李亚飞, 顾铭洪, 程祝宽, 于恒秀. 无选择标记基因的优质香稻新品系培育[J]. 华北农学报, 2015, 30(5): 92-96. doi: 10.7668/hbnxb.2015.05.015.

FENG Meng-shi, YE Jun, HOU Li-li, LI Ming, TANG Ding, ZHANG Jian-xiang, WANG Wei-yun, XU Zhi-hao, LI Ya-fei, GU Ming-hong, CHENG Zhu-kuan, YU Heng-xiu. Breeding of Marker-free Fragrant Rice Via Transgenic Approach[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(5): 92-96. doi: 10.7668/hbnxb.2015.05.015.

http://www.hbnxb.net/CN/Y2015/V30/I5/92

参考文献

[1] 胡培松,唐绍清,顾海华,等.水稻香味的遗传研究与育种利用[J].中国稻米,2006,12(6):1-5.
[2] 于艳敏,王永华.水稻香味研究现状[J].黑龙江农业科学,2010:115-7.
[3] Yajima I,Yanai T,Nakamura M,et al.Volatile flavor components of cooked Kaorimai(Scented Rice,O Sativa Japonica)[J].Agr Biol Chem Tokyo,1979,43:2425-9.
[4] Buttery R G,Ling L C,Juliano B O.2-acetyl-1-pyrroline-an important aroma component of cooked Rice[J].Chem Ind-London,1982:958-959.
[5] Paule C M,Powers J J.Sensory and chemical examination of aromatic and nonaromatic rices[J].J Food Sci,1989,54:343-6.
[6] Bradbury L T,Gillies S A,Brushett D J,et al.Inactivation of an aminoaldehyde dehydrogenase is responsible for fragrance in rice[J].Plant Molecular Biology,2008,68(4/5):439-449.
[7] Thimmaraju R,Bhagyalakshmi N,Narayan M S,et al.In vitro culture of Pandanus amaryllifolius and enhancement of 2-acetyl-1-pyrroline,the major flavouring compound of aromatic rice,by precursor feeding of L-proline[J].Journal of the Science of Food and Agriculture,2005,85(15):2527-2534.
[8] Yoshihashi T,Nguyen T T,Kabaki N.Area dependency of 2-acetyl-1-pyrroline content in an aromatic rice variety,Khao Dawk Mali 105[J].JARQ-Japan Agricultural Research Quarterly,2004,38(2):105-109.
[9] Lin C F,Hsieh T C Y,Hoff B J.Identification and quantification of the popcorn-like aroma in louisiana aromatic della rice(Oryza-Sativa L).[J]Food Sci,1990,55:1466-1467.
[10] Jianming G.Identification of 2-acetylpyridine in Xiangjing- 8618 rice and in Yahonkaoluo leaves[J].Food Chemistry,2002,78(2):163-166.
[11] Chen S,Yang Y,Shi W,et al.Badh2,encoding betaine aldehyde dehydrogenase,inhibits the biosynthesis of 2-acetyl-1-pyrroline,a major component in rice fragrance[J].The Plant Cell,2008,20(7):1850-1861.
[12] Shao G,Tang S,Chen M,et al.Haplotype variation at Badh2, the gene determining fragrance in rice[J].Genomics,2013,101(2):157-162.
[13] Jiamsomboon K,Treesuwan W,Boonyalai N.Dissecting substrate specificity of two rice BADH isoforms:Enzyme kinetics,docking and molecular dynamics simulation studies[J].Biochimie,2012,94(8):1773-1783.
[14] Niu X L,Tang W,Huang W Z,et al.RNAi-directed downregulation of OsBADH2 results in aroma(2-acetyl-1-pyrroline)production in rice(Oryza sativa L.)[J].Bmc Plant Biol,2008,8.
[15] 于恒秀,刘巧泉,王玲,等.根癌农杆菌介导转化水稻成熟胚及转 Metr 基因植株的获得[J].遗传,2005(5):772-778.
[16] Fitzgerald T L,Waters D L,Brooks L O,et al.Fragrance in rice(Oryza sativa)is associated with reduced yield under salt treatment[J].Environmental and Experimental Botany,2010,68(3):292-300.
[17] Bradbury L M T,Fitzgerald T L,Henry R J,et al. The gene for fragrance in rice[J].Plant Biotechnol J,2005,3:363-70.
[18] Yoshihashi T,Huong N T T,Kabaki N.Quality evaluation of Khao Dawk Mali 105,an aromatic rice variety of northeast Thailand[J].JIRCAS Working Report,1999,30:151-60.
[19] Hasthanasombut S,Supaibulwatana K,Mii M,et al.Genetic manipulation of Japonica rice using the OsBADH1 gene from Indica rice to improve salinity tolerance[J].Plant Cell,Tissue and Organ Culture(PCTOC),2011,104(1):79-89.
[20] Shirong J.Safety evaluation of marker genes in transgenic food plants[J].Scientia Agricultura Sinica,1997,2.
[21] Miki B,Mchugh S.Selectable marker genes in transgenic plants:applications,alternatives and biosafety[J].Journal of Biotechnology,2004,107(3):193-232.
[22] Ramessar K,Peremarti A,Gomez-Galera S,et al.Biosafety and risk assessment framework for selectable marker genes in transgenic crop plants:a case of the science not supporting the politics[J].Transgenic Res,2007,16:261-80.
[23] Bai X,Wang Q,Chu C.Excision of a selective marker in transgenic rice using a novel Cre/loxP system controlled by a floral specific promoter[J].Transgenic Research,2008,17(6):1035-1043.
[24] Michelini E,Simoni P,Cevenini L,et al.New trends in bioanalytical tools for the detection of genetically modified organisms:an update[J].Analytical and Bioanalytical Chemistry,2008,392(3):355-367.
[25] Hasthanasombut S,Ntui V,Supaibulwatana K,et al.Expression of indica rice OsBADH1 gene under salinity stress in transgenic tobacco[J].Plant Biotechnology Reports,2010,4(1):75-83.

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无选择标记基因的优质香稻新品系培育

Breeding of Marker-free Fragrant Rice Via Transgenic Approach

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