Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (24): 5248-5260.doi: 10.3864/j.issn.0578-1752.2013.24.019

• RESEARCH NOTES • Previous Articles    

Cloning and Expression Analysis of A Soluble Acid Invertase Gene (SoSAI1) of Sugarcane

 NIU  Jun-Qi-1, WANG  Ai-Qin-1, HUANG  Jing-Li-1, ZHU  Hui-1, LI  Yang-Rui-1, 2 , YANG  Li-Tao-1, 2   

  1. 1.Agricultural College, Guangxi University/State Key Laboratory of Subtropical Bioresources Conservation and Utilization, Nanning 530005;
    2.Sugarcane Research Center, Chinese Academy of Agricultural Sciences/ Sugarcane Research Institute, Guangxi Academy of Agricultural Sciences/ Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture/ Guangxi Key Laboratory of Sugarcane Genetic Improvement, Nanning 530007
  • Received:2013-07-12 Online:2013-12-16 Published:2013-09-27

PDF

1371

Cited

5

Abstract: 【Objective】 The objective of this study is to clone the soluble acid invertase gene (SoSAI1) and 5' promoter sequence from sugarcane, to analyze its sequence and expression patterns. 【Method】 The full-length cDNA sequence of SoSAI1 gene was amplified from the leaves of sugarcane by the technology of rapid amplification of cDNA ends (RACE). The cDNA sequence and the deduced amino acid sequence were analyzed by bioinformatics method. The promoter sequence of SoSAI1 gene was cloned by genome walking. The spatial-specific expressions of SoSAI1 were determined by real-time fluorescent quantitative PCR (qRT-PCR) in leaves and stems at different growth stages of sugarcane, and in roots and leaves under treatment of PEG6000, 100 mmol•L-1 NaCl or 6℃ stress conditions. 【Result】 The cloned full length of SoSAI1 cDNA sequence was 2 387 bp, containing a 2 055 bp open reading frame (ORF) which encodes 685 amino acids with a theoretical molecular mass of 74.44 kD. The 5’ promoter sequence length DNA sequence was 417 bp, which contained endosperm specific expression of cis-acting elements and participate in the drought induced MYB binding sites. The result of real time PCR exhibited that SoSAI1 was the highest abundance in inflorescence and the lowest in old and matured internodes of stems. The conditions of 15% PEG or 6℃ could induce the expression of SoSAI1 gene in leaves, while 15% PEG or NaCl could induce the expression of SoSAI1 gene in roots. 【Conclusion】 The full length of SoSAI1 and part of the promoter were cloned from sugarcane. The analysis by qRT-PCR suggested that SoSAI1 played an important role in sugarcane for growth and development, sucrose accumulation and response to environmental stress.

Key words: sugarcane , soluble acid invertase , gene cloning , expression

[1]Lakshmanan P, Geijskes R J, Aitken K S, Grof C P L, Bonnett G D, Smith G R. Sugarcane biotechnology the challenges and opportunities. In Vitro Cellular and Developmental Biology-Plant, 2005, 41: 345-363.

[2]李杨瑞, 杨丽涛. 20世纪90年代以来我国甘蔗产业和科技的新发展. 西南农业学报, 2009, 22(5): 1469-1475.

Li Y R, Yang L T. New developments in sugarcane industry and technologies in China since 1990s. Southwest China Journal of Agricultual Sciences, 2009, 22(5): 1469-1475. (in Chinese)

[3]Sachdeva M, Bhatia S, Batta S K. Sucrose accumulation in sugarcane: A potential target for crop improvement. Acta Physiology Plant, 2011, 33: 1571-1583.

[4]潘秋红, 张大鹏. 植物转化酶的种类、特性与功能. 植物生理学通讯, 2004, 40(3): 275-280.

Pan Q H, Zhang D P. Isoforms,characteristics and roles of plant invertases. Plant Physiology Communications, 2004, 40(3): 275-280. (in Chinese)

[5]Carson D L, Botha F C. Genes expressed in sugarcane maturing internodal tissue. Plant Cell and Reports, 2002, 20: 1075-1081.

[6]Sturm A. Invertases: Primary structures, functions and roles in plant development and sucrose partitioning. Plant Physiology, 1999, l2l: l-7.

[7]Yamada Y, Kojima T, Bantog N, Shimoda T, Mori H, Shiratake K, Yamaki S. Cloning of two isoforms of soluble acid invertase of Japanese pear and their expression during fruit development. Journal of Plant Physiology, 2007, 164: 746-755.

[8]Tian H M, Kong Q G, Feng Y Q, Yu X Y. Cloning and characterization of a soluble acid invertase-encoding gene from muskmelon. Molecular Biology Reports, 2009, 36: 611-617.

[9]Zhu Y J, Albert H H, Moore P H. Differential expression of soluble acid invertase genes in the shoots of high-sucrose and low-sucrose species of Saccharum and their hybrids. Australian Journal of Plant Physiology, 2000, 27: 193-199.

[10]Sachdeva M, Mann A P S, Batta S K. Sucrose metabolism and expression enzyme activities in low and high storing sugarcane  genotypes of key sucrose. Sugar Technology, 2003, 5(4): 265-271.

[11]潘有强, 罗海玲, 李杨瑞. 甘蔗节间蔗糖含量与蔗糖代谢相关的4种酶活性之间的关系剖析. 植物生理学通讯, 2007, 43(5): 861-864.

Pan Y Q, Luo H L, Li Y R. Correlating annlysis between sucrose content and the activities of four enzymes related to sucrose metabolism in sugarcane(Saccharum officinarum L.) internodes. Plant Physiology Communications, 2007, 43(5): 861-864. (in Chinese)

[12]Pan Y Q, Lou H L, Li Y R. soluble acid invertase and sucrose phosphate synthase: Key enzymes in regulating sucrose accumulation in sugarcane stalk. Sugar Technology, 2009, 11(1): 28-33.

[13]Wang A Q, Huang W J, Niu J Q, Liu M, Yang L T, Li Y R. Effects of ethephon on key enzymes of sucrose metabolism in relation to sucrose accumulation in sugarcane. Sugar Tech, 2013, 15(2): 177-186.

[14]Amin F, Bhatti H N, Asgher M. Partial purification and characterization of an acid invertase from Saccharum Officinarum L.. Pakistan Journal of Botany, 2010, 42(4): 2531-2540.

[15]Hussain A, Rashid M H, Perveen R, Ashraf M. Purification, kinetic and thermodynamic characterization of soluble acid invertase from sugarcane (Saccharum officinarum L.). Plant Physiology and Biochemistry, 2009, 47: 188-194.

[16]Rae A L, Casu R E, Perroux J M, Jackson M A, Grof C P L. A soluble acid invertase is directed to the vacuole by a signal anchor mechanism. Journal of Plant Physiology, 2011, 168: 983-989.

[17]郑月霞, 许玉林, 张积森, 叶冰莹, 陈由强, 陈如凯. 甘蔗可溶性酸性转化酶基因的克隆及序列分析. 热带作物学报, 2012, 33(11): 2007-2011.

Zheng Y X, Xu Y L, Zhang J S, Ye B Y, Chen Y Q, Chen R K. Cloning and sequence analysis of soluble acid invertase from Saccharum officenarum. Chinese Journal of Tropical Crops, 2012, 33(11): 2007-2011. (in Chinese)

[18]刘海斌, 何红, 邓展云, 张革民, 李杨瑞, 方锋学, 贤武, 刘晓静, 方位宽, 闭少玲, 容凤玉, 唐红琴. 特高糖抗寒甘蔗新品种桂糖28的选育. 中国糖料, 2010, 1: 13-15.

Liu H B, He H, Deng Z Y, Zhang G M, Li Y R, Fang F X, Xian B, Liu X J, Fang W K, Bi S L, Rong F Y, Tang H Q. Breeding of new sugarcane variety Guitang 28. Sugar Crops of China, 2010, 1: 13-15. (in Chinese)

[19]应雄美, 蔡青, 毕艳, 刘新龙, 马丽, 毛钧, 陆鑫. 两种不同甘蔗基因组DNA提取方法的比较. 中国糖料, 2009(4): 22-23.

Ying X M, Cai Q, Bi Y, Liu X L, Ma L, Mao J, Lu X. Comparison between two sugarcane genome DNA extraction method. Sugar Crops of China, 2009(4): 22-23. (in Chinese)

[20]阙友雄, 许莉萍, 徐景升, 张积森, 张木清, 陈如凯. 甘蔗基因表达定量PCR分析中内参基因的选择. 热带作物报, 2009, 30(3): 274-278.

Que Y X, Xu L P, Xu J S, Zhang J S, Zhang M Q, Chen R K. Selection of control genes in real-time qPCR analysis of gene expression in sugarcane. Chinese Journal of Tropical Crops, 2009, 30(3): 274-278. (in Chinese)

[21]Iskandar H M, Simpson R S, Casu R E, Bonnett G D, Maclean D J, Manners J M. Comparison of reference genes for quantitative real-time polymerase chain reaction analysis of gene expression in sugarcane. Plant Molecular Biology Reporter, 2004, 22: 325-337.

[22]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt. method. Methods, 2001, 25: 402-408.

[23]Musso M, Bocciardi R, Parodi S, Ravazzolo R, Ceccherini I. Betaine, Dimethyl Sulfoxide, and 7-Deaza-dGTP,a powerful mixture for ampli?cation of GC-rich DNA sequences. Journal of Molecular Diagnostics, 2006, 8: 544-550.

[24]Mammedov T G, Pienaar E, Whitney S E, TerMaat J R, Carvill G, Goliath R, Subramanian A, Viljoen H J. A fundamental study of the PCR amplification of GC-rich DNA templates. Computational Biology and Chemistry, 2008, 32(6): 452-457.

[25]Jensen M A, Fukushima F, Davis R W. DMSO and betaine greatly improve amplification of GC-rich constructs in De Novo synthesis. PLOS One, 2010, 5(6): 1-5.

[26] Hubé F, Reverdiau P, Iochmann S, Gruel Y. Improved PCR method for amplification of GC-Rich DNA sequences. Molecular Biotechnology, 2005, 31(1): 81-85.

[27]Schroeven L, Lammens W, Laere1 A V, Ende1 W V D. Transforming wheat vacuolar invertase into a high affinity sucrose: Sucrose 1-fructosyltransferase. New Phytologist, 2008, 180: 822-831.

[28]Roitsch T, González M C. Function and regulation of plant invertases: Sweet sensations. Trends in Plant Science, 2004, 9(2): 606-613.

[29]Tian H, Kong Q G, Feng Y Q, Yu X Y. Cloning and characterization of a soluble acid invertase-encoding gene from muskmelon. Molecular Biology Reports, 2009, 36: 611-617.

[30]王滕旭, 李正国, 杨迎伍, 邓伟. 哈姆林甜橙蔗糖合酶Ⅰ和酸性转化酶基因表达与果实糖积累的关系. 热带作物学报, 2010, 31(5): 745-749.

Wang T X, Li Z G, Yang Y W, Deng W. Expression of sucrose synthase Ⅰand acid invertase genes in relation to sugar accumulation in fruits of Hamlin sweet orange. Chinese Journal of Tropical Crops, 2010, 31(5): 745-749. (in Chinese)

[31]Verma A K, Upadhyay S K, Srivastava M K, Verma P C, Solomon S, Singh S B. Transcript expression and soluble acid invertase activity during sucrose accumulation in sugarcane. Acta Physiologiae Plantarum, 2011, 33: 1749-1757.

[32]  姚瑞亮, 李杨瑞, 黄玉辉, 杨丽涛, 张桂荣. 甘蔗生长后期乙烯利处理对节间转化酶活性的影响及与蔗糖分积累的关系. 广西农业科学, 2005, 36(2): 106-109.

Yao R L, Li Y R, Huang Y H, Yang L T, Zhang G R. Effects of ethephon on invertases activities inrelation to sucrose accumulation in sugarcane. Guangxi Agricultural Sciences, 2005, 36(2): 106-109. (in Chinese)

[33]Andersen M N, Asch F, Wu Y, Jensen C R, Naested H, Mogensen V O, Koch K E. Soluble invert as expression is an early target of drought stress during the critical, abortion-sensitive phase of young ovary development in maize. Plant Physiology, 2002, 130: 591-604.

[34]Smith A H K, Walker R P, Pollock C J. Invertase in leaves: Conundrum or control point? Journal of Experimental Botany, 1999, 50(335): 735-743.

[35]黄诚梅, 杨翠芳, 潘有强, 魏源文, 邓智年, 吕维莉, 李杨瑞. 可溶性酸性转化酶SAI基因在不同甘蔗基因型中表达分析. 中国糖料, 2013, 2: 21-24.

Huang C M, Yang C F, Pan Y Q, Wei Y W, Deng Z N, Lü W L, Li Y R. Expression analysis of soluble acid invertase gene SAI in different sugarcane (Saccharum officinarum L.) varieties. Sugar Crops of China, 2013, 2: 21-24. (in Chinese)

[36]田晓艳, 刘延吉, 曹敏建. 低钾胁迫对玉米体内N、P、K、转化酶及可溶性糖分配的影响. 玉米科学, 2008, 16(5): 80-82.

Tian X Y, Liu Y J, Cao M J. The distribution of N, P, K, invertase and soluble sugar in maize under low potassium stress. Joumal of Maize Sciences, 2008, 16(5): 80-82. (in Chinese)

[37]姚远, 闵义, 胡新文, 李开绵, 郭建春. 低温胁迫对木薯幼苗叶片转化酶及可溶性糖含量的影响. 热带作物学报, 2010, 31(4): 557-560.

Yao Y, Min Y, Hu X W, Li K M, Guo J C. Effects of low temperature stress on the activity of invertase and soluble sugar content in leaves of cassava seedlings. Chinese Journal of Tropical Crops, 2010, 31(4): 557-560. (in Chinese)

[38]Menossi  M, Silva-Filho M C, Vincentz M, Van-Sluys M A, Souza G M. Sugarcane functional genomics gene discovery for agronomic trait development. International Journal of Plant Genomics, 2008: 1-11.
[1] WANG JiaNuo, CHEN GuiPing, LI Pan, WANG LiPing, NAN YunYou, HE Wei, FAN ZhiLong, HU FaLong, CHAI Qiang, YIN Wen, ZHAO LiaoHao. Photo-Physiological Mechanism at Grain Filling Stage of No-Tillage with Plastic Re-Mulching to Increase Maize Yield in Oasis Irrigation Areas [J]. Scientia Agricultura Sinica, 2026, 59(6): 1189-1202.
[2] CUI ShiYou, CHEN PengJun, MIAO YuanQing, HAN JiJun, SHEN JunMing. Development and Field Evaluation of Glyphosate-Resistant Wheat Germplasm Generated Through EMS Mutagenesis [J]. Scientia Agricultura Sinica, 2026, 59(4): 723-733.
[3] LUO ZhengYing, HU SiZhen, LIN XiuQin, HU Xin, ZHANG Min, XU ChaoHua, LIU XinLong, ZENG QianChun. Identification and Functional Characterization of the PEBP Gene Family in Regulating Flowering Time in Saccharum spontaneum and Saccharum officinarum [J]. Scientia Agricultura Sinica, 2026, 59(4): 734-749.
[4] LIAO TingLu, SHI YaFei, XIAO DongHao, SHE YangMengFei, GUO FuCheng, YANG JiuJu, TANG HaiJiang, LUO ChengKe. The Effect of Exogenous Nitroprusside on Sugar Metabolism in Rice Seedlings Under Alkaline Stress [J]. Scientia Agricultura Sinica, 2026, 59(2): 265-277.
[5] WEI Ping, PAN JuZhong, ZHU DePing, SHAO ShengXue, CHEN ShanShan, WEI YaQian, GAO WeiWei. The Function of OsDREB1J in Regulating Rice Grain Size [J]. Scientia Agricultura Sinica, 2025, 58(8): 1463-1478.
[6] XIONG JiaNi, LI ZongYue, HU HengLiang, GU TianYu, GAO Yan, PENG JiaShi. Influence of Expressing OsNRAMP5 Under the Driving of the OsLCT1 Promoter on Cadmium Migration to Rice Seeds [J]. Scientia Agricultura Sinica, 2025, 58(7): 1259-1268.
[7] YANG CaiLi, LI YongZhou, HE LiangLiang, SONG YinHua, ZHANG Peng, LIU ZhaoXian, LI PengHui, LIU SanJun. Genome-Wide Identification and Analysis of TPS Gene Family and Functional Verification of VvTPS4 in the Formation of Monoterpenes in Grape [J]. Scientia Agricultura Sinica, 2025, 58(7): 1397-1417.
[8] TENG MengXin, XU Ya, HE Jing, WANG Qi, QIAO Fei, LI JingYang, LI XinGuo. Identification and Functional Analysis of Ca2+-ATPase Gene Family in Banana [J]. Scientia Agricultura Sinica, 2025, 58(7): 1418-1433.
[9] PAN LiYuan, WANG YongJun, LI HaiJun, HOU Fu, LI Jing, LI LiLi, SUN SuYang. Screening Regulatory Genes Related to Wheat Grain Protein Accumulation Based on Transcriptome and WGCNA Analysis [J]. Scientia Agricultura Sinica, 2025, 58(6): 1065-1082.
[10] LIU LuPing, HU XueJie, QI Jin, CHEN Qiang, LIU Zhi, ZHAO TianTian, SHI XiaoLei, LIU BingQiang, MENG QingMin, ZHANG MengChen, HAN TianFu, YANG ChunYan. Cloning of the Promoters and Analysis of Expression Patterns of Maturity Genes E1 and E2 in Soybean [J]. Scientia Agricultura Sinica, 2025, 58(5): 840-850.
[11] ZHENG YaQin, LIU XueQing, WU SiWen, TANG XiaoYan, YANG DanNi, WANG YongKang, AHMAD Aftab, KHAN Afrsyab, WANG ChengGang, CHEN GuoHu. Cloning and Expression of BcDET2 Gene and Functional of Its Regulatory Effect on Bolting and Flowering in Wucai (Brassica campestris L.) [J]. Scientia Agricultura Sinica, 2025, 58(5): 991-1003.
[12] ZHANG TianYu, LI Bai, ZANG JinPing, CAO HongZhe, DONG JinGao, XING JiHong, ZHANG Kang. Genome-Wide Identification and Expression Analysis of HMG Family Genes in Botrytis cinerea [J]. Scientia Agricultura Sinica, 2025, 58(4): 704-718.
[13] GUO AoLin, LIN JunXuan, LAI GongTi, HE LiYuan, CHE JianMei, PAN Ruo, YANG FangXue, HUANG YuJi, CHEN GuiXin, LAI ChengChun. Effect of VdF3′5′H2 Overexpression on the Accumulation of Anthocyanin Composition in Spine Grape Cells [J]. Scientia Agricultura Sinica, 2025, 58(4): 802-818.
[14] ZHANG LinLin, GONG Rui, CUI YanLing, ZHONG XiongHui, LI Ye, LI RanHong, QIAN ZongWei. Effect Analysis of SmWRKY30 in Eggplant Resistance to Ralstonia solanacearum by Virus Induced Gene Silencing (VIGS) [J]. Scientia Agricultura Sinica, 2025, 58(3): 548-563.
[15] ZHANG XiangKun, LI JiaYing, QIAO RuMeng, HE JingLei, WANG Li, SHI XiaoXin, DU GuoQiang. Effects of GFabV Under Different Zn Levels on Photosynthetic Efficiency and Photosynthesis-Related Gene Expression of ‘Shine Muscat’ Grapevine [J]. Scientia Agricultura Sinica, 2025, 58(24): 5190-5200.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd