AcCBF1转录因子沉默对低温胁迫下扁桃花器官的影响
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摘要
为了探明AcCBF1基因沉默对‘纸皮’扁桃花器官的影响,以了解CBF转录因子在扁桃花器官发育中的功能,采用病毒诱导的基因沉默(VIGS)技术,以采用农杆菌侵染方法获得的含有pTRV2-AcCBF11、pTRV2-AcCBF12、pTRV2-AcCBF13片段的扁桃AcCBF1基因沉默枝条为试验材料,以常温和低温处理后的自然生长扁桃枝条为对照,观测低温处理后花器官表型指标变化,并对单花药花粉量、花粉活力以及花粉扫描电镜指标结果进行分析。结果表明:低温处理降低了对照组的单花药花粉量和花粉活力,但对花器官指标影响不显著。沉默后的AcCBF12片段和AcCBF13片段显著降低了花粉量与花粉活力,沉默花器官指标效果明显。花粉粒大小无显著变化,花粉纹饰存在一定差异。AcCBF11片段沉默效果不明显。AcCBF12片段和AcCBF13片段影响扁桃花器官表型指标的变化,如花瓣、花冠、花萼、花药、花粉形态、花粉量、花粉活力等,预测CBF转录因子可能在花器官指标发育中发挥着重要功能。
In order to investigate effects of AcCBF1 gene silencing on flower organs in ‘Zhipi' almond, and to learn function of CBF transcription factor during development of almond ?ower organ, using virus induced gene silencing(VIGS) technology, AcCBF1 gene silencing shoots containing pTRV2-AcCBF11, pTRV2-AcCBF12 and pTRV2-AcCBF13 fragments obtained by Agrobacterium inoculation method were used as test materials. After treating at low temperature,changes of phenotypic indexes of ?ower organ were observed, and pollen amount per anther, pollen vigor and pollen SEM results were analyzed. Natural growing almond branches under normal temperature and low temperature treatments were used as controls. The results showed that, low temperature treatment reduced pollen amount per anther and pollen viability in the control group, but had no significant effect on ?ower organ indexs. The silenced AcCBF12 fragment and AcCBF13 fragment significantly reduced pollen amount and pollen viability, and effect of silenced ?ower organ index was obvious. Pollen size did not change significantly, and there were some differences in pollen ornamentation. Silencing effect of AcCBF11 fragment was not obvious. AcCBF12 fragment and AcCBF13 fragment affected phenotypic indexes of almond ?ower organs, such as petal, corolla, calyx, anther, pollen morphology, pollen number, pollen viability, and so on.It was predicted that CBF transcription factor may play an important role during development of ?oral organ indexes.
In order to investigate effects of AcCBF1 gene silencing on flower organs in ‘Zhipi' almond, and to learn function of CBF transcription factor during development of almond ?ower organ, using virus induced gene silencing(VIGS) technology, AcCBF1 gene silencing shoots containing pTRV2-AcCBF11, pTRV2-AcCBF12 and pTRV2-AcCBF13 fragments obtained by Agrobacterium inoculation method were used as test materials. After treating at low temperature,changes of phenotypic indexes of ?ower organ were observed, and pollen amount per anther, pollen vigor and pollen SEM results were analyzed. Natural growing almond branches under normal temperature and low temperature treatments were used as controls. The results showed that, low temperature treatment reduced pollen amount per anther and pollen viability in the control group, but had no significant effect on ?ower organ indexs. The silenced AcCBF12 fragment and AcCBF13 fragment significantly reduced pollen amount and pollen viability, and effect of silenced ?ower organ index was obvious. Pollen size did not change significantly, and there were some differences in pollen ornamentation. Silencing effect of AcCBF11 fragment was not obvious. AcCBF12 fragment and AcCBF13 fragment affected phenotypic indexes of almond ?ower organs, such as petal, corolla, calyx, anther, pollen morphology, pollen number, pollen viability, and so on.It was predicted that CBF transcription factor may play an important role during development of ?oral organ indexes.
引文
[1]李疆.新疆扁桃结实率低的生理与分子机制[J].经济林研究,2011,29(2):1-7.
[2]李鹏,田嘉,唐开文,等.基于隶属函数评估法的扁桃花蕾抗寒性研究[J].中南林业科技大学学报,2017,37(2):39-43.
[3]王敏,李疆,代培红,等.扁桃AcAGL15基因的克隆及植物表达载体构建[J].分子植物育种,2016,14(10):1-9.
[4]BARROS P M,GON?ALVES N,SAIBO N J M,et al.Cold acclimation and floral development in almond bud break:insights into the regulatory pathways[J].Journal of Experimental Botany,2012,63(12):4585-4596.
[5]MARYAM K,ALI E,AMIR M S,et al.Comparison of CBF1,CBF2,CBF3 and CBF4 expression in some grapevine cultivars and species under cold stress[J].Scientia Horticulturae,2015,197:521-526.
[6]YU J,YANG X D,WANG Q,et al.Efficient virus-induced gene silencing in Brassica rapa using a turnip yellow mosaic virus vector[J].Biologia Plantarum,2018,87:1-9.
[7]ZHANG X H,JI N N,MIN D D.A co-silencing system for functional analysis of genes without visible phenotype in tomato plant development and fruit ripening using tobacco rattle virus[J].Scientia Horticulturae,2018,241:100-106.
[8]BOND D M,BAULCOMBE D C.Epigenetic transitions leading to heritable,RNA-mediated de novo silencing in Arabidopsis thaliana[J].Proc Natl Acad Sci USA,2015,112(3):917-922.
[9]LIANG J,CHEN X,ZHAO H,et al.The impacts of BSMVon vegetative growth and water status in hulless barley(Hordeum vulgare var nudum)in VIGS study[J].Acta Societatis Botanicorum Poloniae,2015,84(1):43-51.
[10]JUVALE P S,HEWEZI T,ZHANG C,et al.Temporal and spatial bean pod mottle virus-induced gene silencing in soybean[J].Mol Plant Pathol,2012,13:1140-1148.
[11]TIAN J,CHENG L,HAN Z Y,et al.Tobacco rattle virus mediated gene silencing in strawberry plants[J].Plant Cell,Tissue and Organ Culture,2015,120(3):1131-1138.
[12]GAO X,WHEELER T,LI Z.Silencing GhNDRI and GhMKK2compromises cotton resistance to Verticillium wilt[J].Plant J,2011,66:293-305.
[13]FANTINI E,GIULIANO G.Virus-induced gene silencing as a tool to study tomato fruit biochemistry[J].Plant Signal Transduction,2016,1363:65-78.
[14]JIA H,SHEN Y.Virus-induced gene silencing in strawberry fruit[J].Methods mol Biol,2013,975:211-218.
[15]FOURQUIN C,FERRANDIZ C.Functional analyses of AGAMOUS family members in Nicotiana benthamiana clarify the evolution of early and late roles of C-function genes in eudicots[J].Plant Jounal,2012,71:990-1001.
[16]HSIEH M H,PAN Z J,LAI P H,et al.Virus-induced gene silencing unravels multiple transcription factors involved in floral growth and development in Phalaenopsis orchids[J].Journal of experimental botany,2013,64:3869-3884.
[17]STAMMLER A,MEYER A R,TOWNSLEY B T,et al.Duplicated STM-like KNOX I genes act in floral meristem activity in Eschscholia californica(Papaveraceae)[J].Development genes and evolution,2013,223:289-301.
[18]张洪义.万寿菊(Tagetes erecta)VIGS体系的建立与优化[D].武汉:华中农业大学,2015.
[19]李鹏.扁桃花蕾抗寒性分析和花药开裂过程观察[D].乌鲁木齐:新疆农业大学,2015:33.
[20]李鹏,罗淑萍,李疆.4个栽培扁桃品种和野生扁桃的形态学差异及花粉萌芽率分析[J].塔里木大学学报,2016,28(3):74-79.
[21]田嘉.新疆栽培扁桃花粉形态及授粉亲和性的研究[D].乌鲁木齐:新疆农业大学,2010.
[22]刘晓彬,刘娜,李福宽,等.TRV介导的大豆基因瞬时沉默体系的建立[J].中国农业科学,2015,48(12):2479-2486.
[23]李鹏,田嘉,张琦,等.扁桃离体枝条培养方式比较和筛选[J].中南林业科技大学学报,2018,38(5):23-27.
[24]曲波,翟强,许玉凤.4种忍冬属植物花粉的形态比较[J].沈阳农业大学学报,2006,37(1):96-98.
[25]张锐,毛萌萌,刘振林,等.观赏海棠不同品种花粉形态扫描电镜观察[J].分子植物育种,2018,16(16):5407-5414.
[26]孙琪,廖康,耿文娟,等.基于花粉微观特征的新疆欧洲李亲缘关系分析[J].果树学报,2015,32(3):393-403.
[27]张彦妮,钱灿.12种百合属植物花粉形态扫描电镜观察[J].草业学报,2011,20(5):111-118.
[28]EVRENOSOGLU Y,MISIRLI A.Investigations on the pollen morphology of some fruit species[J].Turk J For Agric,2009,33:181-190.
[29]刘有春,刘威生,刘宁,等.基于花粉形态数量分类的核果类系统关系研究[J].植物遗传资源学报,2010,11(5):645-649.
[30]李燕飞.石斑木属(蔷薇科)植物分类学研究[D].厦门:厦门大学,2014.
[31]王延秀,陈佰鸿,王淑华,等.11个观赏海棠品种花粉形态扫描电镜观察[J].植物研究,2014,34(6):751-757.
[32]梁芳,王冲,宫中志,等.大兴安岭16个地区笃斯越桔花粉形态观察[J].经济林研究,2018,36(2):35-42.
[33]王钦丽,卢龙斗,吴小琴,等.花粉的保存及其生活力测定[J].植物学报,2002,19(3):365-372.
[34]刘鹏,张太奎,王连春,等.牛角瓜花粉离体萌发研究[J].经济林研究,2015,33(2):142-148.
[35]胡琼,王森,郭红艳.不同枣雄株花粉特征和萌发特性[J].经济林研究,2015,33(2):73-81.
[36]刁永强,耿文娟,廖康,等.新疆野生杏花粉量及花粉活力研究初报[J].新疆农业科学,2012,49(1):40-45.
[2]李鹏,田嘉,唐开文,等.基于隶属函数评估法的扁桃花蕾抗寒性研究[J].中南林业科技大学学报,2017,37(2):39-43.
[3]王敏,李疆,代培红,等.扁桃AcAGL15基因的克隆及植物表达载体构建[J].分子植物育种,2016,14(10):1-9.
[4]BARROS P M,GON?ALVES N,SAIBO N J M,et al.Cold acclimation and floral development in almond bud break:insights into the regulatory pathways[J].Journal of Experimental Botany,2012,63(12):4585-4596.
[5]MARYAM K,ALI E,AMIR M S,et al.Comparison of CBF1,CBF2,CBF3 and CBF4 expression in some grapevine cultivars and species under cold stress[J].Scientia Horticulturae,2015,197:521-526.
[6]YU J,YANG X D,WANG Q,et al.Efficient virus-induced gene silencing in Brassica rapa using a turnip yellow mosaic virus vector[J].Biologia Plantarum,2018,87:1-9.
[7]ZHANG X H,JI N N,MIN D D.A co-silencing system for functional analysis of genes without visible phenotype in tomato plant development and fruit ripening using tobacco rattle virus[J].Scientia Horticulturae,2018,241:100-106.
[8]BOND D M,BAULCOMBE D C.Epigenetic transitions leading to heritable,RNA-mediated de novo silencing in Arabidopsis thaliana[J].Proc Natl Acad Sci USA,2015,112(3):917-922.
[9]LIANG J,CHEN X,ZHAO H,et al.The impacts of BSMVon vegetative growth and water status in hulless barley(Hordeum vulgare var nudum)in VIGS study[J].Acta Societatis Botanicorum Poloniae,2015,84(1):43-51.
[10]JUVALE P S,HEWEZI T,ZHANG C,et al.Temporal and spatial bean pod mottle virus-induced gene silencing in soybean[J].Mol Plant Pathol,2012,13:1140-1148.
[11]TIAN J,CHENG L,HAN Z Y,et al.Tobacco rattle virus mediated gene silencing in strawberry plants[J].Plant Cell,Tissue and Organ Culture,2015,120(3):1131-1138.
[12]GAO X,WHEELER T,LI Z.Silencing GhNDRI and GhMKK2compromises cotton resistance to Verticillium wilt[J].Plant J,2011,66:293-305.
[13]FANTINI E,GIULIANO G.Virus-induced gene silencing as a tool to study tomato fruit biochemistry[J].Plant Signal Transduction,2016,1363:65-78.
[14]JIA H,SHEN Y.Virus-induced gene silencing in strawberry fruit[J].Methods mol Biol,2013,975:211-218.
[15]FOURQUIN C,FERRANDIZ C.Functional analyses of AGAMOUS family members in Nicotiana benthamiana clarify the evolution of early and late roles of C-function genes in eudicots[J].Plant Jounal,2012,71:990-1001.
[16]HSIEH M H,PAN Z J,LAI P H,et al.Virus-induced gene silencing unravels multiple transcription factors involved in floral growth and development in Phalaenopsis orchids[J].Journal of experimental botany,2013,64:3869-3884.
[17]STAMMLER A,MEYER A R,TOWNSLEY B T,et al.Duplicated STM-like KNOX I genes act in floral meristem activity in Eschscholia californica(Papaveraceae)[J].Development genes and evolution,2013,223:289-301.
[18]张洪义.万寿菊(Tagetes erecta)VIGS体系的建立与优化[D].武汉:华中农业大学,2015.
[19]李鹏.扁桃花蕾抗寒性分析和花药开裂过程观察[D].乌鲁木齐:新疆农业大学,2015:33.
[20]李鹏,罗淑萍,李疆.4个栽培扁桃品种和野生扁桃的形态学差异及花粉萌芽率分析[J].塔里木大学学报,2016,28(3):74-79.
[21]田嘉.新疆栽培扁桃花粉形态及授粉亲和性的研究[D].乌鲁木齐:新疆农业大学,2010.
[22]刘晓彬,刘娜,李福宽,等.TRV介导的大豆基因瞬时沉默体系的建立[J].中国农业科学,2015,48(12):2479-2486.
[23]李鹏,田嘉,张琦,等.扁桃离体枝条培养方式比较和筛选[J].中南林业科技大学学报,2018,38(5):23-27.
[24]曲波,翟强,许玉凤.4种忍冬属植物花粉的形态比较[J].沈阳农业大学学报,2006,37(1):96-98.
[25]张锐,毛萌萌,刘振林,等.观赏海棠不同品种花粉形态扫描电镜观察[J].分子植物育种,2018,16(16):5407-5414.
[26]孙琪,廖康,耿文娟,等.基于花粉微观特征的新疆欧洲李亲缘关系分析[J].果树学报,2015,32(3):393-403.
[27]张彦妮,钱灿.12种百合属植物花粉形态扫描电镜观察[J].草业学报,2011,20(5):111-118.
[28]EVRENOSOGLU Y,MISIRLI A.Investigations on the pollen morphology of some fruit species[J].Turk J For Agric,2009,33:181-190.
[29]刘有春,刘威生,刘宁,等.基于花粉形态数量分类的核果类系统关系研究[J].植物遗传资源学报,2010,11(5):645-649.
[30]李燕飞.石斑木属(蔷薇科)植物分类学研究[D].厦门:厦门大学,2014.
[31]王延秀,陈佰鸿,王淑华,等.11个观赏海棠品种花粉形态扫描电镜观察[J].植物研究,2014,34(6):751-757.
[32]梁芳,王冲,宫中志,等.大兴安岭16个地区笃斯越桔花粉形态观察[J].经济林研究,2018,36(2):35-42.
[33]王钦丽,卢龙斗,吴小琴,等.花粉的保存及其生活力测定[J].植物学报,2002,19(3):365-372.
[34]刘鹏,张太奎,王连春,等.牛角瓜花粉离体萌发研究[J].经济林研究,2015,33(2):142-148.
[35]胡琼,王森,郭红艳.不同枣雄株花粉特征和萌发特性[J].经济林研究,2015,33(2):73-81.
[36]刁永强,耿文娟,廖康,等.新疆野生杏花粉量及花粉活力研究初报[J].新疆农业科学,2012,49(1):40-45.
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