49种山茶在低温和盐胁迫下的双抗能力分析
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摘要
根据杜鹃红山茶(Camellia azalea)抗坏血酸过氧化物酶(ascorbate peroxidase,APX)基因(CaAPX1,GenBank登录号:KP635267)序列,设计实时荧光定量PCR引物,并对4℃低温和200 mg·L~(-1)NaCl胁迫下杜鹃红山茶CaAPX1表达量进行分析,发现该基因能够在低温和NaCl胁迫下应激上调表达。实时荧光定量PCR分析发现,CaAPX1在49个山茶品种中均得到表达,但表达量有差异。通过田间抗寒耐盐指数和APX1表达量隶属函数值对49个山茶品种抗寒和耐盐双抗能力进行分析,其中Ⅰ类抗寒和耐盐能力强的有4个品种,Ⅱ类抗寒和耐盐能力中等的有11个品种,Ⅲ类抗寒和耐盐能力弱的有34个品种。
On the basic of CaAPX1(GenBank accession ID:KP635267) sequence of Camellia azalea,a pair of qPCR primer was designed and synthesized. Fluorescent quantitative Real-time PCR analysis indicated that the expression levels of Ca APX1 in leaves of Camellia azalea were both upregulated under 4 ℃ and 200 mg · L~(-1) NaCl treatment,reaching peaks at 8 h and 12 h treatment respectively,which indicated that CaAPX1 could be expressed in camellia plants after separately exposed to cold and salt stresses. It was also found that CaAPX1 gene was expressed in the 49 camellia cultivars,but the expression levels were different. Furthermore,on the basis of comprehensive evaluation,49 camellia cultivars were evaluated for their potential of double resistance through the field investigation and CaAPX1 relative expression,the first category(typeⅠ)with strong cold-salt resistance including four camellias;the second category(typeⅡ)to moderate cold-salt resistance including eleven camellias;while the third category(type Ⅲ)with weaker cold-salt resistance including thirty four camellias.
On the basic of CaAPX1(GenBank accession ID:KP635267) sequence of Camellia azalea,a pair of qPCR primer was designed and synthesized. Fluorescent quantitative Real-time PCR analysis indicated that the expression levels of Ca APX1 in leaves of Camellia azalea were both upregulated under 4 ℃ and 200 mg · L~(-1) NaCl treatment,reaching peaks at 8 h and 12 h treatment respectively,which indicated that CaAPX1 could be expressed in camellia plants after separately exposed to cold and salt stresses. It was also found that CaAPX1 gene was expressed in the 49 camellia cultivars,but the expression levels were different. Furthermore,on the basis of comprehensive evaluation,49 camellia cultivars were evaluated for their potential of double resistance through the field investigation and CaAPX1 relative expression,the first category(typeⅠ)with strong cold-salt resistance including four camellias;the second category(typeⅡ)to moderate cold-salt resistance including eleven camellias;while the third category(type Ⅲ)with weaker cold-salt resistance including thirty four camellias.
引文
Chen Dai-hui.2014.The study of cold hardiness physiological and genetic analusis of different Camellias[M.D.Dissertation].Zhengzhou:Henan Agricultural University:1-2.(in Chinese)陈代慧.2014.不同山茶花抗寒生理及遗传差异分析[硕士论文].郑州:河南农业大学:1-2.
Guan Q J,Wang Z J,Wang X H,Takano T,Liu S K.2015.A peroxisomal APX from Puccinellia tenuiflora improves the abiotic stress tolerance of transgenic Arabidopsis thaliana through decreasing of H2O2 accumulation.Journal of Plant Physiology,175:183-191.
Kavitha K,Venkataraman G,Parida A.2008.An oxidative and salinity stress induced peroxisomal ascorbate peroxidase from Avicennia marina:molecular and functional characterization.Plant Physiology and Biochemistry,46:794-804.
Li He-sheng.1992.Principle and technique of experiment in plant physiology and biochemistry.Beijing:Higher Education Press.(in Chinese)李合生.1992.植物生理生化实验原理和技术.北京:高等教育出版社.
Liu Hui-chun.2012.Expression analysis and functional identification of low-temperature-related genes AOX,CAT and APX in Anthurium andraeanum‘Alabama’and construction of genetic transformation system[Ph.D.Dissertation].Hangzhou:Zhejiang University:100-129.(in Chinese)刘慧春.2012.红掌‘阿拉巴马’低温相关基因AOX、CAT和APX的表达分析、功能验证及遗传转化体系的构建[博士论文].杭州:浙江大学:100-129.
Suzuki N,Mittler R.2006.Reactive oxygen species and temperature stress:a delicate balance between signaling and destruction.Physiologia Plantarum,126(1):45-51.
Wang Cai-hua.1988.Fuzzy theory.Beijing:China Architecture Press.(in Chinese)王彩华.1988.模糊论方法学.北京:中国建筑出版社.
Wang J Y,Wu B,Yin H F,Fan Z Q,Li X L,Ni S,He L B,Li J Y.2017.Overexpression of CaAPX induces orchestrated reactive oxygen scavenging and enhances cold and heat tolerances in tobacco.BioMed Research International,doi:org/10.1155/2017/4049534.
Wang Jiang-ying,Shao Xiao-bin,Yin Heng-fu,Fan Zheng-qi,Li Xin-lei,Wu Bin,Ni Sui,Li Ji-yuan.2016.Overexpression of CaAPX1 from Camellia azalea enhances hot stress tolerance in transgenic tobacco.Acta Horticulturae Sinica,43(11):2203-2213.(in Chinese)王江英,邵小斌,殷恒福,范正琪,李辛雷,吴斌,倪穗,李纪元.2016.过量表达杜鹃红山茶CaAPX1的烟草耐热性提高.园艺学报,43(11):2203-2213.
Wang Jiang-ying,Wu Bin,Liu Wei-xin,Fan Zheng-qi,Li Ji-yuan.2015.Evaluation of potential drought-resistance for 24 species in Sect.Theopsis under extreme high temperature and drought in summer.Chinese Journal of Tropical Crops,36(3):563-569.(in Chinese)王江英,吴斌,刘伟鑫,范正琪,李纪元.2015.24种连蕊茶在夏季极端高温干旱条件下的耐寒潜力评价.热带作物学报,36(3):563-569.
Wang Yong-hong.2005.Research on the cold resistance of major Camellia species and some horticultural cultivars[M.D.Dissertation].Beijing:Chinese Academy of Forestry:2-6.(in Chinese)王永红.2005.山茶属主要物种及部分园艺品种耐寒性研究[硕士论文].北京:中国林业科学研究院:2-6.
Xu Chuan-jun,Sun Xu-zhuo,Li Ling,Ru Zhi-wei,Zeng Bi-yu,Liu Yu-mei,Huang Jun-mei.2012.Molecular cloning and expression analysis of homological gene APX from Phalaenopsis.Acta Horticulturae Sinica,39(4):769-776.(in Chinese)许传俊,孙叙卓,李玲,茹志伟,曾碧玉,刘育梅,黄珺梅.2012.蝴蝶兰抗坏血酸过氧化物酶基因克隆及其表达研究.园艺学报,39(4):769-776.
Yang Hua.2007.Eco-physiology responses of Mingshanbaihao to the drought stress[M.D.Dissertation].Ya’an:Sichuan Agricultural University:11-12.(in Chinese)杨华.2007.名山白毫茶树品种对干旱胁迫的生理生态响应[硕士论文].雅安:四川农业大学:11-12.
Zhang Hong-da.1981.Systematic study of Camellia.Journal of Sun Yat-sen University(Natural Science Edition),(1):112.(in Chinese)张宏达.1981.山茶属植物的系统研究.中山大学学报(自然科学版),(1):112.
Guan Q J,Wang Z J,Wang X H,Takano T,Liu S K.2015.A peroxisomal APX from Puccinellia tenuiflora improves the abiotic stress tolerance of transgenic Arabidopsis thaliana through decreasing of H2O2 accumulation.Journal of Plant Physiology,175:183-191.
Kavitha K,Venkataraman G,Parida A.2008.An oxidative and salinity stress induced peroxisomal ascorbate peroxidase from Avicennia marina:molecular and functional characterization.Plant Physiology and Biochemistry,46:794-804.
Li He-sheng.1992.Principle and technique of experiment in plant physiology and biochemistry.Beijing:Higher Education Press.(in Chinese)李合生.1992.植物生理生化实验原理和技术.北京:高等教育出版社.
Liu Hui-chun.2012.Expression analysis and functional identification of low-temperature-related genes AOX,CAT and APX in Anthurium andraeanum‘Alabama’and construction of genetic transformation system[Ph.D.Dissertation].Hangzhou:Zhejiang University:100-129.(in Chinese)刘慧春.2012.红掌‘阿拉巴马’低温相关基因AOX、CAT和APX的表达分析、功能验证及遗传转化体系的构建[博士论文].杭州:浙江大学:100-129.
Suzuki N,Mittler R.2006.Reactive oxygen species and temperature stress:a delicate balance between signaling and destruction.Physiologia Plantarum,126(1):45-51.
Wang Cai-hua.1988.Fuzzy theory.Beijing:China Architecture Press.(in Chinese)王彩华.1988.模糊论方法学.北京:中国建筑出版社.
Wang J Y,Wu B,Yin H F,Fan Z Q,Li X L,Ni S,He L B,Li J Y.2017.Overexpression of CaAPX induces orchestrated reactive oxygen scavenging and enhances cold and heat tolerances in tobacco.BioMed Research International,doi:org/10.1155/2017/4049534.
Wang Jiang-ying,Shao Xiao-bin,Yin Heng-fu,Fan Zheng-qi,Li Xin-lei,Wu Bin,Ni Sui,Li Ji-yuan.2016.Overexpression of CaAPX1 from Camellia azalea enhances hot stress tolerance in transgenic tobacco.Acta Horticulturae Sinica,43(11):2203-2213.(in Chinese)王江英,邵小斌,殷恒福,范正琪,李辛雷,吴斌,倪穗,李纪元.2016.过量表达杜鹃红山茶CaAPX1的烟草耐热性提高.园艺学报,43(11):2203-2213.
Wang Jiang-ying,Wu Bin,Liu Wei-xin,Fan Zheng-qi,Li Ji-yuan.2015.Evaluation of potential drought-resistance for 24 species in Sect.Theopsis under extreme high temperature and drought in summer.Chinese Journal of Tropical Crops,36(3):563-569.(in Chinese)王江英,吴斌,刘伟鑫,范正琪,李纪元.2015.24种连蕊茶在夏季极端高温干旱条件下的耐寒潜力评价.热带作物学报,36(3):563-569.
Wang Yong-hong.2005.Research on the cold resistance of major Camellia species and some horticultural cultivars[M.D.Dissertation].Beijing:Chinese Academy of Forestry:2-6.(in Chinese)王永红.2005.山茶属主要物种及部分园艺品种耐寒性研究[硕士论文].北京:中国林业科学研究院:2-6.
Xu Chuan-jun,Sun Xu-zhuo,Li Ling,Ru Zhi-wei,Zeng Bi-yu,Liu Yu-mei,Huang Jun-mei.2012.Molecular cloning and expression analysis of homological gene APX from Phalaenopsis.Acta Horticulturae Sinica,39(4):769-776.(in Chinese)许传俊,孙叙卓,李玲,茹志伟,曾碧玉,刘育梅,黄珺梅.2012.蝴蝶兰抗坏血酸过氧化物酶基因克隆及其表达研究.园艺学报,39(4):769-776.
Yang Hua.2007.Eco-physiology responses of Mingshanbaihao to the drought stress[M.D.Dissertation].Ya’an:Sichuan Agricultural University:11-12.(in Chinese)杨华.2007.名山白毫茶树品种对干旱胁迫的生理生态响应[硕士论文].雅安:四川农业大学:11-12.
Zhang Hong-da.1981.Systematic study of Camellia.Journal of Sun Yat-sen University(Natural Science Edition),(1):112.(in Chinese)张宏达.1981.山茶属植物的系统研究.中山大学学报(自然科学版),(1):112.