挥发性抑芽物质对马铃薯块茎萌芽的影响及其作用机制
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摘要
马铃薯块茎过早萌芽会降低商品价值,本试验比较挥发性物质的抑芽效应,并从转录和蛋白水平分析其作用机制。结果表明,抑芽能力薄荷醇(29)樟脑(29)萘,萌芽受抑降低代谢消耗,薄荷醇处理180 d时的重量损失只有对照损失的36%。樟脑处理萌芽块茎3 d引起表达显著上调(或下调)的基因和蛋白分别有1227 (299)和296 (204)个,主要参与响应刺激、防御反应。贮藏期间,果胶代谢基因PEL、PME、PG,角质合成基因CYP77A6、HPFT、WES,乙烯合成基因ACO以及转录因子编码基因GATA4L的表达量随时间升高。樟脑早期不同程度地刺激这些基因表达,中后期则抑制,49d时只有对照的0.68%～23.35%。薄荷醇使上述基因表达保持低水平,但可提高细胞周期负控基因KRP4的表达,为对照的15.9倍。植物病原菌互作通路中的基因WRKY75、STH-2、RBOH表达受樟脑诱导并在后期高表达。樟脑和薄荷醇均能抑制生长发育基因的表达,造成芽死亡,降低贮藏损耗。前者早期能促进合成保护性物质,萌芽时有更强烈的抗菌反应;后者则阻碍细胞分裂。
Tuber sprouting losses commercial value. In this experiment, the sprout inhibition abilities of naphthalene, camphor, menthol were researched and the acting mechanism by RNA-seq, iTRAQ was explained. The inhibition abilities showed a trend of menthol > camphor > naphthalene. The metabolic consumption was reduced due to sprouting inhibition and the tuber weight loss of menthol treatment was only 36% of the control loss in 180 days storage. Compared with the control, about 1227(299) and 296(204) genes and proteins whose expression levels were significantly up-regulated(or down-regulated) were detected respectively in sprouting tuber with camphor treatment for three days. Those genes and proteins mainly involved in response to stimulus and defense response. Transcripts of PEL, PME, PG related to pectin degradation, CYP77A6, HPFT, WES related to cutin synthesis, ACO related to synthesis of ethylene, and GATA4 L coding transcription factor were upward with dormancy release during storage. Relative expression of those genes was stimulated at different degrees in camphor treatment at the early stage, but significantly inhibited at the middle and late stages, showing 0.68%–23.35% of the control expression at 49 days. Menthol treatment maintained these genes with low level expression in tuber, but significantly increased the expression of cell cycle inhibitor gene KRP4 to 15.9 times of control. Camphor treatment increased transcripts of genes WRKY75, STH-2, RBOH participating in plant-pathogen interaction pathway to the highest levels at sprouting stage. Therefore, we can conclude that camphor and menthol suppress the growth and development of sprouting tuber, eventually kill the bud and reduce the tuber weight loss during storage. Camphor treatment promotes the biosynthesis of protective substances to resist stress at the early stage and strengthen the resis-tance to pathogen infection at the sprouting stage; menthol treatment prevents bud sprouting by increasing KRP4 expression to inhibit cell division.
Tuber sprouting losses commercial value. In this experiment, the sprout inhibition abilities of naphthalene, camphor, menthol were researched and the acting mechanism by RNA-seq, iTRAQ was explained. The inhibition abilities showed a trend of menthol > camphor > naphthalene. The metabolic consumption was reduced due to sprouting inhibition and the tuber weight loss of menthol treatment was only 36% of the control loss in 180 days storage. Compared with the control, about 1227(299) and 296(204) genes and proteins whose expression levels were significantly up-regulated(or down-regulated) were detected respectively in sprouting tuber with camphor treatment for three days. Those genes and proteins mainly involved in response to stimulus and defense response. Transcripts of PEL, PME, PG related to pectin degradation, CYP77A6, HPFT, WES related to cutin synthesis, ACO related to synthesis of ethylene, and GATA4 L coding transcription factor were upward with dormancy release during storage. Relative expression of those genes was stimulated at different degrees in camphor treatment at the early stage, but significantly inhibited at the middle and late stages, showing 0.68%–23.35% of the control expression at 49 days. Menthol treatment maintained these genes with low level expression in tuber, but significantly increased the expression of cell cycle inhibitor gene KRP4 to 15.9 times of control. Camphor treatment increased transcripts of genes WRKY75, STH-2, RBOH participating in plant-pathogen interaction pathway to the highest levels at sprouting stage. Therefore, we can conclude that camphor and menthol suppress the growth and development of sprouting tuber, eventually kill the bud and reduce the tuber weight loss during storage. Camphor treatment promotes the biosynthesis of protective substances to resist stress at the early stage and strengthen the resis-tance to pathogen infection at the sprouting stage; menthol treatment prevents bud sprouting by increasing KRP4 expression to inhibit cell division.
引文
[1]肖关丽,郭华春.不同生理年龄马铃薯种薯芽中的内源激素含量变化及其对马铃薯植株生长发育的影响.植物生理学报,2007,43:818-820.Xiao G L,Guo H C.Changes in endogenous hormone contents in bud of seed potato(Solanum tuberosum L.)with different physiological ages and its effect on growth and development.Plant Physiol J,2007,43:818-820(in Chinese with English abstract).
[2]Daniels-Lake B J,Pruski K,Prange R K.Using ethylene gas and chlorpropham potato sprout inhibitors together.Potato Res,2011,54:223-236.
[3]Vijay P,Ezekiel R,Pandey R.Use of CIPC as a potato sprout suppressant:health and environmental concerns and future options.Qual Assur Safety Crops Foods,2018,10:17-24.
[4]Gómez-Castillo D.Effects of essential oils on sprout suppression and quality of potato cultivars.Postharvest Biol Technol,2013,82:15-21.
[5]Hartmans K J.The use of carvone in agriculture:sprout suppression of potatoes and antifungal activity against potato tuber and other plant diseases.Ind Crops Prod,1995,4:3-13.
[6]Oosterhaven K,Poolman B,Smid E J.S-carvone as a natural potato sprout inhibiting,fungistatic and bacteristatic compound.Ind Crops Prod,1995,4:23-31.
[7]Teper-Bamnolker P,Dudai N,Fischer R,Belausov E,Zemach H,Shoseyov O,Eshel D.Mint essential oil can induce or inhibit potato sprouing by differential alteration of apical meristem.Planta,2010,232:179-186.
[8]Weerd J W,Thornton M K,Shafii B.Sprout suppressing residue levels of 1,4-dimethylnaphthalene(1,4DMN)in potato cultivars.Am J Potato Res,2010,87:434-445.
[9]Campbell M A,Gleichsner A,Alsbury R,Horvath D,Suttle J.The sprout inhibitors chlorpropham and 1,4-dimethylnaphthalene elicit different transcriptional profiles and do not suppress growth through a prolongation of the dormant state.Plant Mol Biol,2010,73:181-189.
[10]Campbell M A,Gleichsner A,Hilldorfer L,Horvath D,Suttle J.The sprout inhibitor 1,4-dimethylnaphthalene induces the expression of the cell cycle inhibitors KRP1 and KRP2 in potatoes.Funct Integr Genomics,2012,12:533-541.
[11]Li L Q,Zou X,Deng M S,Peng J,Huang X L,Lu X,Fang C C,Wang X Y.Comparative morphology,transcription,and proteomics study revealing the key molecular mechanism of camphor on the potato tuber sprouting effect.Int J Mol Sci,2017,18:2280.
[12]Liu B L,Zhang N,Wen Y K,Jin X,Yang J W,Si H J,Wang D.Transcriptomic changes during tuber dormancy release process revealed by RNA sequencing in potato.J Biotechnol,2015,198:17-30.
[13]Yang Y,Qiang X,Owsiany K,Zhang S,Thannhauser T W,Li L.Evaluation of different multidimensional LC-MS/MS pipelines for isobaric tags for relative and absolute quantitation(iTRAQ)-based proteomic analysis of potato tubers in response to cold storage.J Proteome Res,2011,10:4647-4660.
[14]The Potato Genome Sequencing Consortium.Genome sequence and analysis of the tuber crop potato.Nature,2011,475:189-195.
[15]Mortazavi A,Williams B A,McCue K,Schaeffer L,Wold B.Mapping and quantifying mammalian transcriptomes by RNA-Seq.Nat Methods,2008,5:621-628.
[16]邹雪,邓孟胜,李立芹,余金龙,丁凡,黄雪丽,彭洁,帅禹,蔡诚诚,王西瑶.油菜素内酯合成和信号转导基因在马铃薯块茎贮藏期间的表达变化及对萌芽的影响.作物学报,2017,43:811-820.Zou X,Deng M S,Li L Q,Yu J L,Ding F,Huang X L,Peng J,Shuai Y,Cai C C,Wang X Y.Expression changes of genes related to brassinosteroid biosynthesis and signal transduction during potato storage and its effect on tuber sprouting.Acta Agron Sin,2017,43:811-820(in Chinese with English abstract).
[17]Dixon R A.Natural products and plant disease resistance.Nature,2001,411:843-847.
[18]Kobayashi M,Ohura I,Kawakita K,Yokota N,Fujiwara M,Shimamoto K,Doke N,Yoshioka H.Calcium-dependent protein kinases regulate the production of reactive oxygen species by potato NADPH oxidase.Plant Cell,2007,19:1065-1080.
[19]Baydar H,Karado?an T.The effects of volatile oils on in vitro potato sprout growth.Potato Res,2003,46:1-8.
[20]马进,郑钢,裴翠明,张振亚.基于iTRAQ质谱分析技术筛选南方型紫花苜蓿根部响应盐胁迫差异表达蛋白.农业生物技术学报,2016,24:497-509.Ma J,Zheng G,Pei C M,Zhang Z Y.Screening differentially expressed proteins in southern type alfalfa(Medicago sativa‘Millenium’)root upon salt stress by iTRAQ protein mass spectrometry.J Agric Biotechnol,2016,24:497-509(in Chinese with English abstract).
[21]Takahashi K,Niwa H,Yokota N,Kubota K,Inoue H.Widespread tissue expression of nepenthesis-like aspartic protease genes in Arabidopsis thaliana.Plant Physiol Biol,2008,46:724-729.
[22]李合生.现代植物生理学(第3版).北京:高等教育出版社,2012.pp 245-246.Li H S.Modern Plant Physiology,3rd edn.Beijing:Higher Education Press,2012.pp 245-246(in Chinese).
[23]Coleman W K,Dioxide C.Oxygen and ethylene effects on potato tuber dormancy release and sprout growth.Ann Bot,1998,82:21-27
[24]Hartmann A,Senning M,Hedden P,Sonnewald U,Sonnewald S.Reactivation of meristem activity and sprout growth in potato tubers require both cytokinin and gibberellin.Plant Physiol,2011,155:776-796.
[25]Külen O,Stushnoff C,Davidson R D,Holm D G.Gibberelllic acid and ethephon alter potato minituber bud dormancy and improve seed tuber yield.Am J Potato Res,2011,88:167-174.
[26]Amita C,Sane V A,Nath P.Differential expression of pectate lyase during ethylene-induced postharvest softening of mango(Mangifera indica var.Dashehari).Physiol Planta,2006,128:546-555.
[27]Singh A P,Pandey S P,Rajluxmi,Pandey S,Nath P,Sane A P.Transcriptional activation of a pectate lyase gene,RbPel1,during petal abscission in rose.Postharvest Biol Technol,2011,60:143-148.
[28]Ranftl Q L,Bastakis E,Klermund C,Schwechheimer C.LLM-domain containing B-GATA factors control different aspects of cytokinin-regulated development in Arabidopsis thaliana.Plant Physiol,2016,170:2295-2311.
[29]Luo X M,Lin W H,Zhu S W,Zhu J Y,Sun Y,Fan X Y,Cheng ML,Hao Y Q,Oh E,Tian M M,Liu L J,Zhang M,Xie Q,Chong K,Wang Z Y.Integration of light-and brassinosteroid-signaling pathways by a GATA transcription factor in Arabidopsis.Dev Cell,2010,19:872-883.
[30]Shin J M,Chung K M,Sakamoto S,Kojima S,Yeh C M,Ikeda M,Mitsuda N,Ohme-Takagi M.The chimeric repressor for the GATA4 transcription factor improves tolerance to nitrogen deficiency in Arabidopsis.Plant Biotechnol,2017,34:151-158.
[31]Leene J V,Hollunder J,Eeckhout D,Persiau G,Slijke E V D,Stals H.Targeted interactomics reveals a complex core cell cycle machinery in Arabidopsis thaliana.Mol Sys Biol,2010,6:397.
[32]Sang E J,Yoko O,Jaesung N,Masaaki U,Gyung-Tae K.Kip-related protein 3 is required for control of endoreduplication in the shoot apical meristem and leaves of Arabidopsis.Mol Cells,2013,35:47-53.
[33]Cheng Y,Cao L,Wang S,Li Y P,Shi X Z.Downregulation of multiple CDK inhibitor ICK/KRP genes upregulates the E2Fpathway and increases cell proliferation,and organ and seed sizes in Arabidopsis.Plant J,2013,75:642-655.
[34]Constabel C P,Brisson N.The defense-related STH-2 gene product of potato shows race-specific accumulation after inoculation with low concentrations of Phytophthora infestans zoospores.Planta,1992,188:289-295.
[35]Constabel C P,Bertrand C,Brisson N.Transgenic potato plants overexpressing the pathogenesis-related STH-2 gene show unaltered susceptibility to Phytophthora infestans and potato virus X.Plant Mol Biol,1993,22:775-782.
[36]El-Banna A,Hajirezaei M R,Wissing J,Ali Z,Vaas L,Heine-Dobbernack E,Jacobsen H J,Schumacher H M,Kiesecker H.Over-expression of PR-10a leads to increased salt and osmotic tolerance in potato cell cultures.J Biotechnol,2010,150:277-287.
[37]Park S,Gupta R,Krishna R,Kim S T,Lee D Y,Hwang D J,Bae S C,Ahn I P.Proteome analysis of disease resistance against Ralstonia solanacearum in potato cultivar CT206-10.Plant Pathol J,2016,32:25-32.
[2]Daniels-Lake B J,Pruski K,Prange R K.Using ethylene gas and chlorpropham potato sprout inhibitors together.Potato Res,2011,54:223-236.
[3]Vijay P,Ezekiel R,Pandey R.Use of CIPC as a potato sprout suppressant:health and environmental concerns and future options.Qual Assur Safety Crops Foods,2018,10:17-24.
[4]Gómez-Castillo D.Effects of essential oils on sprout suppression and quality of potato cultivars.Postharvest Biol Technol,2013,82:15-21.
[5]Hartmans K J.The use of carvone in agriculture:sprout suppression of potatoes and antifungal activity against potato tuber and other plant diseases.Ind Crops Prod,1995,4:3-13.
[6]Oosterhaven K,Poolman B,Smid E J.S-carvone as a natural potato sprout inhibiting,fungistatic and bacteristatic compound.Ind Crops Prod,1995,4:23-31.
[7]Teper-Bamnolker P,Dudai N,Fischer R,Belausov E,Zemach H,Shoseyov O,Eshel D.Mint essential oil can induce or inhibit potato sprouing by differential alteration of apical meristem.Planta,2010,232:179-186.
[8]Weerd J W,Thornton M K,Shafii B.Sprout suppressing residue levels of 1,4-dimethylnaphthalene(1,4DMN)in potato cultivars.Am J Potato Res,2010,87:434-445.
[9]Campbell M A,Gleichsner A,Alsbury R,Horvath D,Suttle J.The sprout inhibitors chlorpropham and 1,4-dimethylnaphthalene elicit different transcriptional profiles and do not suppress growth through a prolongation of the dormant state.Plant Mol Biol,2010,73:181-189.
[10]Campbell M A,Gleichsner A,Hilldorfer L,Horvath D,Suttle J.The sprout inhibitor 1,4-dimethylnaphthalene induces the expression of the cell cycle inhibitors KRP1 and KRP2 in potatoes.Funct Integr Genomics,2012,12:533-541.
[11]Li L Q,Zou X,Deng M S,Peng J,Huang X L,Lu X,Fang C C,Wang X Y.Comparative morphology,transcription,and proteomics study revealing the key molecular mechanism of camphor on the potato tuber sprouting effect.Int J Mol Sci,2017,18:2280.
[12]Liu B L,Zhang N,Wen Y K,Jin X,Yang J W,Si H J,Wang D.Transcriptomic changes during tuber dormancy release process revealed by RNA sequencing in potato.J Biotechnol,2015,198:17-30.
[13]Yang Y,Qiang X,Owsiany K,Zhang S,Thannhauser T W,Li L.Evaluation of different multidimensional LC-MS/MS pipelines for isobaric tags for relative and absolute quantitation(iTRAQ)-based proteomic analysis of potato tubers in response to cold storage.J Proteome Res,2011,10:4647-4660.
[14]The Potato Genome Sequencing Consortium.Genome sequence and analysis of the tuber crop potato.Nature,2011,475:189-195.
[15]Mortazavi A,Williams B A,McCue K,Schaeffer L,Wold B.Mapping and quantifying mammalian transcriptomes by RNA-Seq.Nat Methods,2008,5:621-628.
[16]邹雪,邓孟胜,李立芹,余金龙,丁凡,黄雪丽,彭洁,帅禹,蔡诚诚,王西瑶.油菜素内酯合成和信号转导基因在马铃薯块茎贮藏期间的表达变化及对萌芽的影响.作物学报,2017,43:811-820.Zou X,Deng M S,Li L Q,Yu J L,Ding F,Huang X L,Peng J,Shuai Y,Cai C C,Wang X Y.Expression changes of genes related to brassinosteroid biosynthesis and signal transduction during potato storage and its effect on tuber sprouting.Acta Agron Sin,2017,43:811-820(in Chinese with English abstract).
[17]Dixon R A.Natural products and plant disease resistance.Nature,2001,411:843-847.
[18]Kobayashi M,Ohura I,Kawakita K,Yokota N,Fujiwara M,Shimamoto K,Doke N,Yoshioka H.Calcium-dependent protein kinases regulate the production of reactive oxygen species by potato NADPH oxidase.Plant Cell,2007,19:1065-1080.
[19]Baydar H,Karado?an T.The effects of volatile oils on in vitro potato sprout growth.Potato Res,2003,46:1-8.
[20]马进,郑钢,裴翠明,张振亚.基于iTRAQ质谱分析技术筛选南方型紫花苜蓿根部响应盐胁迫差异表达蛋白.农业生物技术学报,2016,24:497-509.Ma J,Zheng G,Pei C M,Zhang Z Y.Screening differentially expressed proteins in southern type alfalfa(Medicago sativa‘Millenium’)root upon salt stress by iTRAQ protein mass spectrometry.J Agric Biotechnol,2016,24:497-509(in Chinese with English abstract).
[21]Takahashi K,Niwa H,Yokota N,Kubota K,Inoue H.Widespread tissue expression of nepenthesis-like aspartic protease genes in Arabidopsis thaliana.Plant Physiol Biol,2008,46:724-729.
[22]李合生.现代植物生理学(第3版).北京:高等教育出版社,2012.pp 245-246.Li H S.Modern Plant Physiology,3rd edn.Beijing:Higher Education Press,2012.pp 245-246(in Chinese).
[23]Coleman W K,Dioxide C.Oxygen and ethylene effects on potato tuber dormancy release and sprout growth.Ann Bot,1998,82:21-27
[24]Hartmann A,Senning M,Hedden P,Sonnewald U,Sonnewald S.Reactivation of meristem activity and sprout growth in potato tubers require both cytokinin and gibberellin.Plant Physiol,2011,155:776-796.
[25]Külen O,Stushnoff C,Davidson R D,Holm D G.Gibberelllic acid and ethephon alter potato minituber bud dormancy and improve seed tuber yield.Am J Potato Res,2011,88:167-174.
[26]Amita C,Sane V A,Nath P.Differential expression of pectate lyase during ethylene-induced postharvest softening of mango(Mangifera indica var.Dashehari).Physiol Planta,2006,128:546-555.
[27]Singh A P,Pandey S P,Rajluxmi,Pandey S,Nath P,Sane A P.Transcriptional activation of a pectate lyase gene,RbPel1,during petal abscission in rose.Postharvest Biol Technol,2011,60:143-148.
[28]Ranftl Q L,Bastakis E,Klermund C,Schwechheimer C.LLM-domain containing B-GATA factors control different aspects of cytokinin-regulated development in Arabidopsis thaliana.Plant Physiol,2016,170:2295-2311.
[29]Luo X M,Lin W H,Zhu S W,Zhu J Y,Sun Y,Fan X Y,Cheng ML,Hao Y Q,Oh E,Tian M M,Liu L J,Zhang M,Xie Q,Chong K,Wang Z Y.Integration of light-and brassinosteroid-signaling pathways by a GATA transcription factor in Arabidopsis.Dev Cell,2010,19:872-883.
[30]Shin J M,Chung K M,Sakamoto S,Kojima S,Yeh C M,Ikeda M,Mitsuda N,Ohme-Takagi M.The chimeric repressor for the GATA4 transcription factor improves tolerance to nitrogen deficiency in Arabidopsis.Plant Biotechnol,2017,34:151-158.
[31]Leene J V,Hollunder J,Eeckhout D,Persiau G,Slijke E V D,Stals H.Targeted interactomics reveals a complex core cell cycle machinery in Arabidopsis thaliana.Mol Sys Biol,2010,6:397.
[32]Sang E J,Yoko O,Jaesung N,Masaaki U,Gyung-Tae K.Kip-related protein 3 is required for control of endoreduplication in the shoot apical meristem and leaves of Arabidopsis.Mol Cells,2013,35:47-53.
[33]Cheng Y,Cao L,Wang S,Li Y P,Shi X Z.Downregulation of multiple CDK inhibitor ICK/KRP genes upregulates the E2Fpathway and increases cell proliferation,and organ and seed sizes in Arabidopsis.Plant J,2013,75:642-655.
[34]Constabel C P,Brisson N.The defense-related STH-2 gene product of potato shows race-specific accumulation after inoculation with low concentrations of Phytophthora infestans zoospores.Planta,1992,188:289-295.
[35]Constabel C P,Bertrand C,Brisson N.Transgenic potato plants overexpressing the pathogenesis-related STH-2 gene show unaltered susceptibility to Phytophthora infestans and potato virus X.Plant Mol Biol,1993,22:775-782.
[36]El-Banna A,Hajirezaei M R,Wissing J,Ali Z,Vaas L,Heine-Dobbernack E,Jacobsen H J,Schumacher H M,Kiesecker H.Over-expression of PR-10a leads to increased salt and osmotic tolerance in potato cell cultures.J Biotechnol,2010,150:277-287.
[37]Park S,Gupta R,Krishna R,Kim S T,Lee D Y,Hwang D J,Bae S C,Ahn I P.Proteome analysis of disease resistance against Ralstonia solanacearum in potato cultivar CT206-10.Plant Pathol J,2016,32:25-32.