CAT activity and related gene expression in the pistil of self- and cross-pollinated Camellia oleifera Abel.
-
摘要: 油茶(Camellia oleifera Abel.)具有后期自交不亲和(LSI)特性，严重阻碍了油茶杂交育种的亲本选择和品种配置。为研究过氧化氢酶(Catalase, CAT)在油茶自交不亲和中的作用，本文对油茶自、异交授粉后雌蕊中的CAT活性进行了测定，并克隆得到3个CAT基因，命名为CoCAT1～CoCAT3。结果显示，其CDS分别为1479、1287和1479 bp，编码492、428和492个氨基酸。多序列比对结果表明，其编码产物在C末端均含有典型的三肽序列。系统进化分析发现，CoCAT2、CoCAT3与NtCAT3亲缘关系最近，CoCAT1与CsCAT1亲缘关系最近。基因表达模式分析结果显示，3个基因在授粉72 h后的自交雌蕊中表达量最大，在授粉36、84 h自交雌蕊中的表达量显著高于异交。自、异花授粉均引起油茶雌蕊内CAT活性的上升，但幅度不同，推测油茶CAT可能参与油茶自交不亲和的响应机制。Abstract: Camellia oleifera Abel. shows late-acting self-incompatibility (LSI) characteristics,which seriously hinder parental selection and cultivar arrangement during cross breeding. To study the role of catalase (CAT) in the self-incompatibility of C. oleifera, CAT activities in the pistils after self- and cross-pollination were determined. Three CAT genes (CoCAT1, CoCAT2, and CoCAT3) were cloned by reverse-transcription polymerase chain reaction (RT-PCR), with coding sequences (CDSs)of 1479, 1287, and 1479 bp, encoding 492, 428, and 492 amino acids, respectively. Multiple sequence alignment showed that their coding products contained typical tripeptide sequences at the C-terminal.Phylogenetic analysis showed that CoCAT2 and CoCAT3 were most closely related to NtCAT3, and CoCAT1 was most closely related to CsCAT1. Expression pattern analysis indicated that the expression levels of the three genes were highest in the self-pollinated pistils 72 h after pollination, and expression levels in the self-pollinated pistils at 36 and 84 h were significantly higher than those in the cross-pollinated pistils.Thus, self-pollination and cross-pollination increased CAT activity in the pistils of C. oleifera, but at different magnitudes. This study indicated that CAT maybe involved in the response mechanism of self-incompatibility in C. oleifera.
-
Keywords:
- Camellia oleifera /
- Self-incompatibility /
- Catalase /
- Gene cloning /
- Expression pattern
-
-
[1] 曹彦妮. 油茶FBPase基因和GAPDH基因的全长cDNA克隆及原核表达分析[D]. 长沙:中南林业科技大学, 2011:1-71. [2] 高超, 杨瑞, 郭其强, 袁德义. 油茶柱头和花柱的显微与超微结构特征[J]. 林业科学研究, 2019, 32(1):1-7. Gao C, Yang R, Guo QQ, Yuan DY. Microstructure and ultrastructure characteristics of stigma and style of Camellia oleifera[J]. Forest Research, 2019, 32(1):1-7.
[3] 廖婷, 袁德义, 彭邵锋, 邹锋. 油茶自交和异交过程花粉管生长的荧光显微观察[J]. 中南林业科技大学学报, 2012, 32(7):34-37. Liao T, Yuan DY, Peng SF, Zou F.A fluorescence microscope observation on self and cross-pollination of pollen tubes in Camellia oleifer a[J]. Journal of Central South University of Forestry & Technology, 2012, 32(7):34-37.
[4] 高超. 油茶后期自交不亲和性的细胞学研究[D]. 长沙:中南林业科技大学, 2017:1-107. [5] Liao T, Yuan DY, Zou F, Gao C, Yang Y, et al. Self-steri-lity in Camellia oleifera may be due to the prezygotic late-acting self-incompatibility[J]. PLoS One, 2014, 9(6):e99639.
[6] 廖婷. 油茶自交不亲和性初步研究[D]. 长沙:中南林业科技大学, 2013:1-75. [7] Ogata M. Acatalasemia[J]. Hum Genet, 1991, 86(4):331-340.
[8] 陈卫国, 刘勇, 周冀衡. 低温胁迫对烟草膜保护酶系统的影响[J]. 安徽农业科学, 2011, 39(7):3978-3980. Chen WG, Liu Y, Zhou JH. Influence of low temperature stress on the membrane-protected enzyme system in tobacco[J]. Journal of Anhui Agricultural Sciences, 2011, 39(7):3978-3980.
[9] 贾晨晨. 葡萄过氧化氢酶基因在毕赤酵母中的表达及酶学特性的初级研究[D]. 南京:南京农业大学, 2012:1-94. [10] Willekens H, Chamnongpol S, Davey M, Schraudner M, Langebartels C, et al. Catalase is a sink for H2O2 and is indispensable for stress defence in C3 plants[J]. EMBO J, 1997, 16(16):4806-4816.
[11] 宋新华, 赵凤云. 植物体内过氧化氢酶的研究进展[J]. 安徽农业科学, 2007, 35(31):9824-9827. Song XH, Zhao FY. Research progress on catalase in plants[J]. Journal of Anhui Agricultural Sciences, 2007, 35(31):9824-9827.
[12] Li J, Liu JT, Wang GQ, Cha JY, Li GN, et al. A chape-rone function of NO CATALASE ACTIVITY1 is required to maintain catalase activity and for multiple stress responses in Arabidopsis[J]. Plant Cell, 2015, 27(3):908-925.
[13] Li Q, Yang A, Zhang WH. Comparative studies on tole-rance of rice genotypes differing in their tolerance to mo-derate salt stress[J]. BMC Plant Biol, 2017, 17(1):141.
[14] Redinbaugh MG, Wadsworth GJ, Scandalios JG. Characterization of catalase transcripts and their differential expression in maize[J]. Biochim Biophys Acta (BBA) -Gene Struct Expression, 1988, 951(1):104-116.
[15] 侯含, 王升平, 张超群, 周志成, 余祥文, 等. 烟草过氧化氢酶基因CAT2克隆与表达特征分析[J]. 中国烟草科学, 2019, 40(1):1-8. Hou H, Wang SP, Zhang CQ, Zhou ZC, Yu XW, et al. Cloning of Catalase 2(CAT2) gene and study on its expression pattern in Nicotiana tabacum L.[J]. Chinese Tobacco Science, 2019, 40(1):1-8.
[16] 林星谷, 孔德仓, 庞晓明, 李颖岳. 冬枣过氧化氢酶基因的克隆及表达分析[J].西北植物学报, 2012, 32(6):1086-1092. Lin XG, Kong DC, Pang XM, Li YY. Cloning and tissue expression analysis of catalase gene in Dongzao (Zi-ziphus jujuba)[J]. Acta Botanica Boreali-Occidentalia Sinica, 2012, 32(6):1086-1092.
[17] 吴能表, 徐光德, 唐于婷, 朱利泉, 王小佳. 自交不亲和甘蓝的花粉萌发与花柱内保护酶活性变化[J]. 西南师范大学学报(自然科学版), 2004, 29(5):848-851. Wu NB, Xu GD, Tang YT, Zhu LQ, Wang XJ. Pollen germination and change of protective enzyme activity in style of self-incompatible Brassica oleracea L.[J]. Journal of Southwest China Normal University (Natural Science Edition), 2004, 29(5):848-851.
[18] 王保成, 孙万仓, 范惠玲, 孟亚雄, 马静芳,等. 芸芥自交亲和系与自交不亲和系SOD、POD和CAT酶活性[J]. 中国油料作物学报, 2006, 28(2):162-165, 171. Wang BC, Sun WC, Fan HL, Meng YX, Ma JF, et al. Studies on SOD、POD and CAT activity between self-compatible and self-incompatible lines in Eruca sativa Mill[J]. Chinese Journal of Oil Crop Sciences, 2006, 28(2):162-165, 171.
[19] 张雪梅, 李保国, 赵志磊, 郭素萍, 齐国辉. 苹果自花授粉花粉管生长和花柱保护酶活性与内源激素含量的关系[J]. 林业科学, 2009, 45(11):20-25. Zhang XM, Li BG, Zhao ZL, Guo SP, Qi GH. Relationship between compatibility of self-pollination and changes in protecting enzyme and hormone in different apple cultivars[J]. Scientia Silvae Sinicae, 2009, 45(11):20-25.
[20] Zhou JQ, Lu MQ, Yu SS, Liu YY,Yang J, Tan XF. In-depth understanding of Camellia oleifera self-incompatibility by comparative transcriptome, proteome and metabolome[J]. Int J Mol Sci, 2020, 21(5):1600.
[21] 谭晓风, 袁德义, 袁军, 邹锋, 谢鹏, 等. 大果油茶良种'华硕'[J]. 林业科学, 2011, 47(12):184, 209. Tan XF, Yuan DY, Yuan J, Zou F, Xie P, et al.An elite variety:Camellia oleifera 'Huashuo'[J]. Scientia Silvae Sinicae, 2011, 47(12):184, 209.
[22] 袁德义, 谭晓风, 邹锋, 袁军, 谢鹏, 等. 油茶良种'华金'[J]. 林业科学, 2012, 48(2):187, 190. Yuan DY, Tan XF, Zou F, Yuan J, Xie P, et al.An elite variety:Camellia oleifera 'Huajin'[J]. Scientia Silvae Sinicae, 2012, 48(2):187, 190.
[23] 王群, 刘朝巍, 徐文娟. 紫外分光光度法测定玉米过氧化氢酶活性新进展[J]. 中国农学通报, 2016, 32(15):159-165. Wang Q, Liu CW, Xu WJ. Ultraviolet spectrophotometry measurement of catalase activity in maize[J]. Chinese Agricultural Science Bulletin, 2016, 32(15):159-165.
[24] Marshall OJ. PerlPrimer:cross-platform, graphical primer design for standard, bisulphite and real-time PCR[J]. Bioinformatics, 2004, 20(15):2471-2472.
[25] Xiao XL, Ma JB, Wang JR, Wu XM, Li PB, Yao YN. Validation of suitable reference genes for gene expression analysis in the halophyte Salicornia europaea by real-time quantitative PCR[J]. Front Plant Sci, 2015, 5:788.
[26] Loewen PC, Villanueva J, Switala J, Donald LJ, Ivancich A. Unprecedented access of phenolic substrates to the heme active site of a catalase:substrate binding and peroxidase-like reactivity of Bacillus pumilus catalase monitored by X-ray crystallography and EPR spectroscopy[J]. Proteins Struct Funct Bioinf, 2015, 83(5):853-866.
[27] Zafra A, Rodríguez-García MI, de Dios Alché J. Cellular localization of ROS and NO in olive reproductive tissues during flower development[J]. BMC Plant Biol, 2010, 10:36.
[28] Wilkins KA, Bancroft J, Bosch M, Ings J, Smirnoff N, Franklin-Tong VE. Reactive oxygen species and nitric oxide mediate actin reorganization and programmed cell death in the self-incompatibility response of Papaver[J]. Plant Physiol, 2011, 156(1):404-416.
[29] Wheeler MJ, de Graaf BHJ, Hadjiosif N, Perry RM, Poulter NS, et al. Identification of the pollen self-incompatibility determinant in Papaver rhoeas[J]. Nature, 2009, 459(7249):992-995.
[30] Sewelam N, Kazan K, Schenk PM. Global plant stress signaling:reactive oxygen species at the cross-road[J]. Front Plant Sci, 2016, 7:187.
[31] Yang T, Poovaiah BW. Hydrogen peroxide homeostasis:activation of plant catalase by calcium/calmodulin[J]. Proc Natl Acad Sci USA, 2002, 99(6):4097-4102.
[32] 熊博, 汪志辉, 廖凤玲, 范茜茜, 李立佼, 王晓晓. 钙对黄果柑果实3种抗氧化酶活性的影响[J]. 植物科学学报, 2014, 32(2):168-173. Xiong B, Wang ZH, Liao FL, Fan QQ, Li LJ, Wang XX. Effects of Calcium on the activities of three antioxidant enzymes in Huangguogan fruit[J]. Plant Science Journal, 2014, 32(2):168-173.
[33] Dat JF, Pellinen R, Beeckman T, Van De Cotte B, Langebartels C, et al. Changes in hydrogen peroxide homeostasis trigger an active cell death process in tobacco[J]. Plant J, 2003, 33(4):621-632.
[34] 向警, 黄倩, 鞠春燕, 黄伦霄,赵正武. 外源褪黑素对盐胁迫下水稻种子萌发与幼苗生长的影响[J]. 植物生理学报, 2021, 57(2):393-401. Xiang J, Huang Q, Ju CY, Huang LX, Zhao ZW. Effect of exogenous melatonin on seed germination and seedling growth of rice under salt stress[J]. Plant Physiology Journal, 2021, 57(2):393-401.
[35] 吕芳, 丁刚, 詹冬梅, 王翔宇, 吴海一. 茉莉酸甲酯对铜藻生长、抗氧化系统及岩藻黄素含量的影响[J]. 植物生理学报, 2019, 55(5):667-675. Lü F, Ding G, Zhan DM, Wang XY, Wu HY. Effect of methyl jasmonate on the growth, antioxidant system and fucoxanthin content of Sargassum horneri[J]. Plant Physio-logy Journal, 2019, 55(5):667-675.
[36] 黄玲, 赖佳, 韦树谷, 代顺冬, 李琼英, 等. 石刁柏两性花与雄花的抗氧化酶活性及其相关基因可变剪接的差异分析[J]. 植物生理学报, 2019, 55(8):1231-1238. Huang L, Lai J, Wei SG, Dai SD, Li QY, et al. Diffe-rences of antioxidant enzyme activities and alternative splicing of related genes between hermaphroditic and male flower buds in asparagus[J]. Plant Physiology Journal, 2019, 55(8):1231-1238.
[37] 范晓明, 袁德义, 李建中, 田晓明, 熊欢, 朱周俊. 锥栗花序分化发育过程中的生理生化变化[J]. 植物生理学报, 2017, 53(12):2206-2214. Fan XM, Yuan DY, Li JZ, Tian XM, Xiong H, Zhu ZJ. Physiological and biochemical changes in Castanea henryi during the inflorescence differentiation[J]. Plant Physiology Journal, 2017, 53(12):2206-2214.
[38] 高超, 袁德义, 杨亚, 王碧芳, 刘冬明, 等. 油茶自交不亲和性的解剖特征[J]. 林业科学, 2015, 51(2):60-68. Gao C, Yuan DY, Yang Y, Wang BF, Liu DM, et al. Anatomical characteristics of self-Incompatibility in Camellia oleifera[J]. Scientia Silvae Sinicae, 2015, 51(2):60-68.
[39] 李欢,陈莹, 黑小斌, 张娜, 张岗, 郭顺星. 铁皮石斛过氧化氢酶基因DoCAT1的鉴定与表达分析[J]. 中国药学杂志, 2018, 53(17):1447-1452. Li H, Chen Y, Hei XB, Zhang N, Zhang G, Guo SX. Identification and expression analysis of a catalase gene DoCAT1 in Dendrobiu mofficinale[J]. Chinese Pharmaceutical Journal, 2018, 53(17):1447-1452.
计量
- 文章访问数: 125
- HTML全文浏览量: 1
- PDF下载量: 22
下载:
