比克氏棉色素腺体形态建成与棉酚代谢特性的研究
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摘要
【目的】对比克氏棉(G.bickii Prokh)种子形成过程和种子萌发过程中的色素腺体形态建成特点、色素腺体与棉酚含量的动态关系及其分子机理进行探究。【方法】以比克氏棉为材料,进行组织切片观察、棉酚含量测定并分析对种子萌发期色素腺体形成及棉酚合成基因的表达谱。【结果】比克氏棉在种胚形成过程中,子叶上形成大量色素腺体原,并在种子萌发后12 h形成腔体结构,转换成成熟的色素腺体;比克氏棉种胚形成过程及种子萌发过程中均未检测到棉酚,但从种子萌发12 h开始可以检测到较低含量的棉酚。种子萌发期色素腺体形成及棉酚合成相关基因表达谱分析结果表明,GoPGF基因在比克氏棉种子萌发12 h的子叶上表达量较高,与比克氏棉色素腺体形成有关;法尼基焦磷酸合成酶(FPS)基因在种子萌发24 h的子叶上表达量较高,可能与棉酚向子叶色素腺体转运有关;CYP706B1基因在种子萌发0 h较高,HMG1基因在种子萌发0 h、36 h和48 h的子叶上表达量均较高,说明这两个基因与种子萌发后色素腺体腔的形成有关,并参与棉酚的贮运;杜松烯合酶基因(CAD1-A)和HMG2基因与比克氏棉色素腺体形成无密切关系。【结论】种子萌发后的12 h为比克氏棉子叶色素腺体形态建成的关键时期,且色素腺体的形态建成与棉酚出现的时间具有同步性。GoPGF、FPS、CYP706B1、HMG1参与色素腺体的形成及棉酚的转运。
[Objective]We studied the process of pigment gland formation and the dynamic change of gossypol synthesis,the dynamic relationship and molecular mechanism between the pigment glands and gossypol during the process of embryo formation and seed germination in G.bickii Prokh.[Method] We chose G.bickii as meterial and studied the tissue section observation,determination of gossypol contents and gene expression profile analysis of pigment gland formation and gossypol synthesis genes.[Result] The results showed that a large number of pigment protoglands formed on the cotyledons during embryo development process in G.bickii.The pigment protoglands formed to cavity structure at 12 hours after seed germination,and then converted into mature pigment glands.Analysis of gene expression profiles related to the pigment gland formation and gossypol synthesis during seed germination showing that Go PGF gene was highly expressed on the cotyledons of seed germination of 12 hours,which may be related to pigment protoglands forming to cavity structure of G.bickii.Farnesyl pyrophosphate synthase gene(FPS) was highly expressed on the cotyledons of seed germination of 24 hours showing it may be associated with the gossypol transported into the cotyledon pigment glands.CYP706 B1 gene was highly expressed on the cotyledons of seed germination of 0 hours and HMG1 were highly expressed on the cotyledons of seed germination of 0 hours,36 hours and 48 hours showing these genes may be associated with the formation of pigmented cavities and the storage and transport of gossypol;Cadinene synthase genes(CAD1-A) and HMG2 genes were not closely related to the pigment gland formation.[Conclusion] The 12 hours after seed germination is the key period of the morphogenesis of the pigment glands,and the morphogenesis of the pigment glands is synchrony with the time of the occurrence of gossypol in G.bickii.GoPGF,FPS,CYP706 B1 and HMG1 are associated with the formation of pigmented cavities and the storage and transport of gossypol.
[Objective]We studied the process of pigment gland formation and the dynamic change of gossypol synthesis,the dynamic relationship and molecular mechanism between the pigment glands and gossypol during the process of embryo formation and seed germination in G.bickii Prokh.[Method] We chose G.bickii as meterial and studied the tissue section observation,determination of gossypol contents and gene expression profile analysis of pigment gland formation and gossypol synthesis genes.[Result] The results showed that a large number of pigment protoglands formed on the cotyledons during embryo development process in G.bickii.The pigment protoglands formed to cavity structure at 12 hours after seed germination,and then converted into mature pigment glands.Analysis of gene expression profiles related to the pigment gland formation and gossypol synthesis during seed germination showing that Go PGF gene was highly expressed on the cotyledons of seed germination of 12 hours,which may be related to pigment protoglands forming to cavity structure of G.bickii.Farnesyl pyrophosphate synthase gene(FPS) was highly expressed on the cotyledons of seed germination of 24 hours showing it may be associated with the gossypol transported into the cotyledon pigment glands.CYP706 B1 gene was highly expressed on the cotyledons of seed germination of 0 hours and HMG1 were highly expressed on the cotyledons of seed germination of 0 hours,36 hours and 48 hours showing these genes may be associated with the formation of pigmented cavities and the storage and transport of gossypol;Cadinene synthase genes(CAD1-A) and HMG2 genes were not closely related to the pigment gland formation.[Conclusion] The 12 hours after seed germination is the key period of the morphogenesis of the pigment glands,and the morphogenesis of the pigment glands is synchrony with the time of the occurrence of gossypol in G.bickii.GoPGF,FPS,CYP706 B1 and HMG1 are associated with the formation of pigmented cavities and the storage and transport of gossypol.
引文
[1]刘毓湘,吴忠道,于绍杰,等.无腺体棉育种与棉籽利用[M].北京:中国农业出版社,1988:1-22.Liu Yuxiang,Wu Zhongdao,Yu Shaojie,et al.Breeding of glandless cotton and utilization of cottonseed[M].Beijing:China Agricultural Press,1988:1-22.
[2]Sunilkumar G,Campbell L M,Puckhaber L,et al.Engineering cottonseed for use in human nutrition by tissue-specific reduction of toxic gossypol[J].Proceedings of the National Academy of Sciences of the United States of America,2006,103(48):18054-18059.
[3]朱四元,陈金湘.棉酚腺体的遗传分析及低酚棉分子育种研究进展[J].中国农学通报,2005,21(9):57-60.Zhu Siyuan,Chen Jinxiang.Genetic analysis of pigment glands and progress in molecular breeding of low phenolic cotton[J].Chinese Agricultural Science Bulletin,2005,21(9):57-60.
[4]祝水金,季道藩,刘胜安,等.棉花色素腺体和棉酚对棉铃虫生长发育和抗药性的影响[J].中国农业科学,2001,34(2):157-162.Zhu Shuijin,Ji Daofan,Liu Sheng'an,et al.Effects of cotton pigment glands and gossypol on the growth,development and resistance of cotton bollworm[J].Scientia Agricultura Sinica,2001,34(2):157-162.
[5]钱玉源,张海娜,崔淑芳,等.棉花色素腺体的研究进展[J].作物研究,2017,31(2):206-209.Qian Yuyuan,Zhang Haina,Cui Shufang,et al.Progress in the study of cotton pigment glands[J].Crop Research,2017,31(2):206-209.
[6]Liu Wenzhe,Zhou Yafu,Wang Xin,et al.Programmed cell death during pigment gland formation in Gossypium hirsutum leaves[J].Plant Biology,2010,12(6):895-902.
[7]祝水金,蒋玉蓉,Reddy N,等.陆地棉子叶色素腺体延缓形成种质系的育成及其遗研究[J].科学通报,2004,49(19):1987-1992.Zhu Shuijin,Jiang Yurong,Reddy N,et al.Development and study of a germplasm line for delayed formation of pigment glands in upland cotton[J].Chinese Science Bulletin,2004,49(19):1987-1992.
[8]童旭宏,祝水金,季道藩.陆地棉子叶色素腺体延缓形成性状的表现与组织结构观察[J].棉花学报,2005,17(3):137-140.Tong Xuhong,Zhu Shuijin,Ji Daofan.The Expression and anatomical observation of the delayed pigment gland morphogenesis in an upland cotton germplasm[J].Cotton Science,200517(3):137-140.
[9]Punit M,Singh P,Dongre A B,et al.Variability for gossypol glands in upland cotton(Gossypium hirsutum)[J].Advances in Plant Sciences,1991,4(1):165-170.
[10]Punit M,Singh I D.Growth and infestation of boll weevils on normal-glanded,glandless,and high-gossypol strains of cotton[J].Journal of Economic Entomology,1972,3(65):821-824.
[11]丁亮,祝水金,胡丹艳,等.比克氏棉和司笃克氏棉色素腺体形态建成的组织结构观察[J].棉花学报,2003,15(1):17-21.Ding Liang,Zhu Shuijin,Hu Danyan,et al.Observation on the anatomical structure of pigment glands in G.bickii and G.stockii[J].Cotton Science,2003,15(1):17-21.
[12]Leandro L L,Harlan C S,Norma L T,et al.Hmg-coA reductase gene family in cotton(Gossypium hirsutum L.):Unique structural features and differential expression of hmg2 potentially associated with synthesis of specific isoprenoids in developing embryos[J].Plant and Cell Physiology,1999,40(7):750-761.
[13]Ma Dan,Hu Yan,Yang Changqing,et al.Genetic basis for glandular trichome formation in cotton[J].Nature Communications2016,7:10456.http:/doi.org/10.103/ncomms10456.
[14]毛晓霞.石蜡切片制作方法的改良[J].安徽农学通报,201319(8):15-17.Mao Xiaoxia.Improvement of the method of making paraffin section[J].Anhui Agricultural Science Bulletin,2013,19(8):15-17.
[15]刘慧,季灵艳,赵天伦,等.陆地棉色素腺体与不同棉酚旋光体含量之间的相关性研究[J].棉花学报,2017,29(5):437-446.Liu Hui,Ji Lingyan,Zhao Tianlun,et al.Analysis of the correlation between pigment glands and(-)/(+)-gossypol contents in upland cotton[J].Cotton Science,2017,29(5):437-446.
[16]祝水金,季道藩.澳洲野生棉种子叶色素腺体延缓形成性状的遗传研究[J].科学通报,2011,46(2):132-137.Zhu Shuijin,Ji Daofan.Genetic study on the delayed formation of wild cotton pigment glands in Australian wild cotton[J].Chinese Science Bulletin,2011,46(2):132-137.
[17]项时康,吴也文,褚志亮,等.陆地棉幼苗棉酚动态分析[J].中国棉花,1988,15(3):20-22.Xiang Shikang,Wu Yewen,Zhu Zhiliang,et al.Dynamic analysis of gossypol in the seedlings of upland cotton[J].China Cotton,1988,15(3):20-22.
[18]胡丹艳.棉花色素腺体的形态结构与转育利用研究[D].杭州:浙江大学,2002.Hu Danyan.Studies on the anatomocal structures and utilization in cotton breeding for the pigment gland[D].Hangzhou:Zhejiang University,2002.
[19]邱新棉.试论低酚棉资源及其利用前景[J].中国棉花,2000,27(2):7-9.Qiu Xinmian.Low phenol cotton resources and utilization prospects[J].China Cotton,2000,27(2):7-9.
[20]李炳林.种子无腺体植株有腺体棉花异源四倍体新种质的育成和研究[J].棉花学报,1993,3(1):27-32.Li Binglin.Breeding a new germplasm of glandless plant with glandless cotton heterologous tetraploid[J].Cotton Science,1993,3(1):27-32.
[21]张雪林,金林,张天真,等.种子低酚植株有酚的棉花新品种[J].中国棉花,1984,11(6):19-21.Zhang Xuelin,Jin Lin,Zhang Tianzhen,et al.A new cotton variety with phenolics in low phenol plants[J].China Cotton,1984,11(6):19-21.
[22]汪若海.我国低酚棉育种的进展[J].中国棉花,1990,17(2):4-6.Wang Ruohai.Progress in breeding of low phenolic cotton in China[J].China Cotton,1990,17(2):4-6.
[23]袁有禄,靖深蓉,张天真,等.陆地棉显性无腺体指示性状的杂种优势利用研究[J].种子,1997,2(6):15-17.Yuan Youlu,Jing Shenrong,Zhang Tianzhen,et al.Study on heterosis utilization of dominant glandless indicator characters in upland cotton[J].Seed,1997,2(6):15-17.
[24]钱玉源,刘祎,张海娜,等.棉酚合成及棉花腺体形成相关基因的研究进展[J].棉花学报,2017,29(3):301-306.Qian Yuyuan,Liu Yi,Zhang Haina,et al.Advances in genes related to gossypol biosynthesis and cotton gland formation[J].Cotton Science,2017,29(3):301-306.
[25]Berardi L C.Toxic constituents of plant foodstuffs[M].New York:Academic Press,1980:213-363.
[2]Sunilkumar G,Campbell L M,Puckhaber L,et al.Engineering cottonseed for use in human nutrition by tissue-specific reduction of toxic gossypol[J].Proceedings of the National Academy of Sciences of the United States of America,2006,103(48):18054-18059.
[3]朱四元,陈金湘.棉酚腺体的遗传分析及低酚棉分子育种研究进展[J].中国农学通报,2005,21(9):57-60.Zhu Siyuan,Chen Jinxiang.Genetic analysis of pigment glands and progress in molecular breeding of low phenolic cotton[J].Chinese Agricultural Science Bulletin,2005,21(9):57-60.
[4]祝水金,季道藩,刘胜安,等.棉花色素腺体和棉酚对棉铃虫生长发育和抗药性的影响[J].中国农业科学,2001,34(2):157-162.Zhu Shuijin,Ji Daofan,Liu Sheng'an,et al.Effects of cotton pigment glands and gossypol on the growth,development and resistance of cotton bollworm[J].Scientia Agricultura Sinica,2001,34(2):157-162.
[5]钱玉源,张海娜,崔淑芳,等.棉花色素腺体的研究进展[J].作物研究,2017,31(2):206-209.Qian Yuyuan,Zhang Haina,Cui Shufang,et al.Progress in the study of cotton pigment glands[J].Crop Research,2017,31(2):206-209.
[6]Liu Wenzhe,Zhou Yafu,Wang Xin,et al.Programmed cell death during pigment gland formation in Gossypium hirsutum leaves[J].Plant Biology,2010,12(6):895-902.
[7]祝水金,蒋玉蓉,Reddy N,等.陆地棉子叶色素腺体延缓形成种质系的育成及其遗研究[J].科学通报,2004,49(19):1987-1992.Zhu Shuijin,Jiang Yurong,Reddy N,et al.Development and study of a germplasm line for delayed formation of pigment glands in upland cotton[J].Chinese Science Bulletin,2004,49(19):1987-1992.
[8]童旭宏,祝水金,季道藩.陆地棉子叶色素腺体延缓形成性状的表现与组织结构观察[J].棉花学报,2005,17(3):137-140.Tong Xuhong,Zhu Shuijin,Ji Daofan.The Expression and anatomical observation of the delayed pigment gland morphogenesis in an upland cotton germplasm[J].Cotton Science,200517(3):137-140.
[9]Punit M,Singh P,Dongre A B,et al.Variability for gossypol glands in upland cotton(Gossypium hirsutum)[J].Advances in Plant Sciences,1991,4(1):165-170.
[10]Punit M,Singh I D.Growth and infestation of boll weevils on normal-glanded,glandless,and high-gossypol strains of cotton[J].Journal of Economic Entomology,1972,3(65):821-824.
[11]丁亮,祝水金,胡丹艳,等.比克氏棉和司笃克氏棉色素腺体形态建成的组织结构观察[J].棉花学报,2003,15(1):17-21.Ding Liang,Zhu Shuijin,Hu Danyan,et al.Observation on the anatomical structure of pigment glands in G.bickii and G.stockii[J].Cotton Science,2003,15(1):17-21.
[12]Leandro L L,Harlan C S,Norma L T,et al.Hmg-coA reductase gene family in cotton(Gossypium hirsutum L.):Unique structural features and differential expression of hmg2 potentially associated with synthesis of specific isoprenoids in developing embryos[J].Plant and Cell Physiology,1999,40(7):750-761.
[13]Ma Dan,Hu Yan,Yang Changqing,et al.Genetic basis for glandular trichome formation in cotton[J].Nature Communications2016,7:10456.http:/doi.org/10.103/ncomms10456.
[14]毛晓霞.石蜡切片制作方法的改良[J].安徽农学通报,201319(8):15-17.Mao Xiaoxia.Improvement of the method of making paraffin section[J].Anhui Agricultural Science Bulletin,2013,19(8):15-17.
[15]刘慧,季灵艳,赵天伦,等.陆地棉色素腺体与不同棉酚旋光体含量之间的相关性研究[J].棉花学报,2017,29(5):437-446.Liu Hui,Ji Lingyan,Zhao Tianlun,et al.Analysis of the correlation between pigment glands and(-)/(+)-gossypol contents in upland cotton[J].Cotton Science,2017,29(5):437-446.
[16]祝水金,季道藩.澳洲野生棉种子叶色素腺体延缓形成性状的遗传研究[J].科学通报,2011,46(2):132-137.Zhu Shuijin,Ji Daofan.Genetic study on the delayed formation of wild cotton pigment glands in Australian wild cotton[J].Chinese Science Bulletin,2011,46(2):132-137.
[17]项时康,吴也文,褚志亮,等.陆地棉幼苗棉酚动态分析[J].中国棉花,1988,15(3):20-22.Xiang Shikang,Wu Yewen,Zhu Zhiliang,et al.Dynamic analysis of gossypol in the seedlings of upland cotton[J].China Cotton,1988,15(3):20-22.
[18]胡丹艳.棉花色素腺体的形态结构与转育利用研究[D].杭州:浙江大学,2002.Hu Danyan.Studies on the anatomocal structures and utilization in cotton breeding for the pigment gland[D].Hangzhou:Zhejiang University,2002.
[19]邱新棉.试论低酚棉资源及其利用前景[J].中国棉花,2000,27(2):7-9.Qiu Xinmian.Low phenol cotton resources and utilization prospects[J].China Cotton,2000,27(2):7-9.
[20]李炳林.种子无腺体植株有腺体棉花异源四倍体新种质的育成和研究[J].棉花学报,1993,3(1):27-32.Li Binglin.Breeding a new germplasm of glandless plant with glandless cotton heterologous tetraploid[J].Cotton Science,1993,3(1):27-32.
[21]张雪林,金林,张天真,等.种子低酚植株有酚的棉花新品种[J].中国棉花,1984,11(6):19-21.Zhang Xuelin,Jin Lin,Zhang Tianzhen,et al.A new cotton variety with phenolics in low phenol plants[J].China Cotton,1984,11(6):19-21.
[22]汪若海.我国低酚棉育种的进展[J].中国棉花,1990,17(2):4-6.Wang Ruohai.Progress in breeding of low phenolic cotton in China[J].China Cotton,1990,17(2):4-6.
[23]袁有禄,靖深蓉,张天真,等.陆地棉显性无腺体指示性状的杂种优势利用研究[J].种子,1997,2(6):15-17.Yuan Youlu,Jing Shenrong,Zhang Tianzhen,et al.Study on heterosis utilization of dominant glandless indicator characters in upland cotton[J].Seed,1997,2(6):15-17.
[24]钱玉源,刘祎,张海娜,等.棉酚合成及棉花腺体形成相关基因的研究进展[J].棉花学报,2017,29(3):301-306.Qian Yuyuan,Liu Yi,Zhang Haina,et al.Advances in genes related to gossypol biosynthesis and cotton gland formation[J].Cotton Science,2017,29(3):301-306.
[25]Berardi L C.Toxic constituents of plant foodstuffs[M].New York:Academic Press,1980:213-363.