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| Invertase Activity and Related Genes Expression Analysis in Cytoplasmic Male Sterile and Fertile Pepper (Capsicum annuum) Lines |
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Abstract Sucrose is one of the main assimilates in plant photosynthesis process. The effective use of sucrose in plant reproduction plays important roles in the process of fertilization and fruit-setting. Invertase (INV), as a key enzyme of sucrose metabolism, regulate the hydrolysis of sucrose into glucose and fructose. According to the cellular localization, INVs are further divided as cytoplasmic invertase (CIN), vacuolar invertase (VIN), and cell wall invertase (CWIN). In order to understand the physiological and molecular mechanism of sucrose metabolism involved in the process of male sterility of pepper (Capsicum annuum) induced by pollen abortion, the INV enzyme activities, INV genes expression pattern and their correlation with pepper male sterility were analyzed. A pair of pepper male sterile line 131A and maintainer line 131B (fertile) were used as investigating materials in this study. The parameters including INV enzyme activities, VIN and CWIN gene identification in pepper genome, and CWIN encoding gene expression pattern by qRT-PCR, were comparatively analyzed in different tissues and developmental stages of pepper flower. The results showed that the CWIN activities in the big bud and flower of the maintainer line 131B were significantly higher than those in the male sterile line 131A (P＜0.001 and P＜0.01, respectively), which were closely correlated with the high expression level of pepper CaCWIN encoding genes CaCWIN3 and CaCWIN4. According to the results, CaCWIN3 and CaCWIN4 were likely the key functional genes involved in the process of male sterility of pepper. These results shed light on the further investigation of molecular regulation mechanism of capsicum male sterility.
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Received: 26 April 2018
Published: 20 November 2018
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| [1]戴雄泽, 唐冬英.辣椒雄性不育与花蕾内源激素含量变化的关系[J].辣椒杂志, 2004, (1):10-13[2]郭爽, 常绍东, 黄贞, 等.辣椒雄性不育研究进展[J].辣椒杂志, 2012, (2):1-5[3]刘科伟, 王述彬, 刘金兵, 等.辣椒细胞质雄性不育基因的AFLP分析[J].分子植物育种, 2009, 6:58-60[4]阮美颖, 李志邈, 程远, 等.单性结实与非单性结实樱桃番茄的蔗糖转化酶与相关基因的表达分析[J].农业与生物技术学报, 2016, 24(2):206-214[5]孙日飞, 方智远, 张淑江, 等.萝卜胞质大白菜雄性不育系的生化分析[J].园艺学报, 2000, (3):187-192[6]邹学校, 侯喜林, 刘荣云, 等.辣椒细胞质雄性不育基因对不育系及杂交一代农艺性状和生化特性的影响.[J].园艺学报, 2004, (6):732-736[7]Barratt D H, Derbyshire P, Findlay K, et al.Normal growth of Arabidopsis requires cytosolic invertase but not sucrose synthase[J].Proceedings of the National Academy of Sciences of the USA, 2009, 106(31):13124-13129[8]Bradford M.A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biochemistry, 1976, 72:248-254[9]Fridman E, Zamir D.Functional divergence of a syntenic invertase gene family in tomato, potato, and Arabidopsis[J].Plant Physiology, 2003, 131(2):603-609[10]Jia L, Zhang B, Mao C, et al.OsCYT-INV1 for alkaline/ neutral invertase is involved in root cell development and reproductivity in rice (Oryza sativa L.)[J].Planta, 2008, 228(1):51-59[11]Jin Y, Ni D A, Ruan Y L.Posttranslational elevation of cell wall invertase activity by silencing its inhibitor in tomato delays leaf senescence and increases seed weight and fruit hexose level[J].Plant Cell, 2009, 21(7):2072-2089[12]Letunic I, Doerks T, Bork P, et al.SMART 7: recent updates to the protein domain annotation resource[J].Nucleic Acids Res, 2012, 40:D302-D305[13]Livak K J, and Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method [J].Methods, 2001, 25:402-408[14]Li Z, Palmer W M, Martin A P, et al.High invertase activity in tomato reproductive organs correlates with enhanced sucrose import into, and heat tolerance of, young fruit[J].Journal of Experimental Botany, 2012, 63(3):1155-1166[15]Punta M, Coggill P C, Eberhardt R Y, et al.The Pfam protein families database[J].IEEE International Conference on Acoustics, 2009, :809-812[16]Qin C, Yu C, Shen Y, et al.Whole-genome sequencing of cultivated and wild peppers provides insights into Capsicum domestication and specialization[J].Proceedings of the National Academy of Sciences , USA, 2014, 111(14):5135-5140[17]Ruan Y L.Sucrose metabolism: Gateway to diverse carbon use and sugar signaling[J].Annual Review of Plant Biology, 2014, 65:33-67[18]Ruan Y L, Jin Y, Yang Y J, et al.Sugar input, metabolism, and signaling mediated by invertase: Roles in development, yield potential, and response to drought and heat[J].Molecular Plant, 2010, 3(6):942-955[19]Sturm A.Invertases. Primary structures, functions, and roles in plant development and sucrose partitioning[J].Plant Physiology, 1999, 121(1):1-8[20]Tamura K, Peterson D, Peterson N, et al.MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J].Molecular Biology & Evolution, 2011, 28:2731-2739[21]Thompson J D, Gibson T J, Plewniak F, et al.The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research, 1997, 25:4876-4882[22]Tomlinson K L, McHugh S, Labbe H, et al.Evidence that the hexose-to-sucrose ratio does not control the switch to storage product accumulation in oilseeds: Analysis of tobacco seed development and effects of overexpressing apoplastic invertase[J].Journal of Experimental Botany, 2004, 55(406):2291-2303[23]Wan H J, Yuan W, Ruan M Y, et al.Identification of reference genes for reverse transcription quantitative real-time PCR normalization in pepper (Capsicum annuum L.)[J].Biochemical & Biophysical Research Communication, 2011, 416:24-30[24]Wang L, Li X R, Lian H, et al.Evidence that high activity of vacuolar invertase is required for cotton fiber and Arabidopsis root elongation through osmotic dependent and independent pathways, respectively[J].Plant Physiology, 2010, 154(2):744-756[25]Zanor MI, Osorio S, Nunes-Nesi A, et al.RNA interference of LIN5 in Solanum lycopersicum confirms its role in controlling Brix content, uncovers the influence of sugars on the levels of fruit hormones and demonstrates the importance of sucrose cleavage for normal fruit development and fertility[J].Plant Physiology, 2009, 150(3):1204-1218 |
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