Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (7): 1387-1396.doi: 10.3864/j.issn.0578-1752.2014.07.016

• HORTICULTURE • Previous Articles     Next Articles

Cloning of a Plasma Membrane Aquaporin Gene EjPIP1 in Eriobotrya japonica Leaves and Its Expression Analysis After AM Fungi Inoculation

 ZHANG  Yan-1, 2 , LI  Juan-3, YAO  Qing-1, CHEN  Jie-Zhong-1, HU  You-Li-1   

  1. 1、College of Horticulture, South China Agricultural University, Guangzhou 510642；
    2、Agricultural College，Guizhou University/Guizhou Engineering Research Center for Fruit Crops，Guiyang 550025；
    3、Zhongkai University of Agriculture and Engineering，Guangzhou 510225
  • Received:2013-07-10 Online:2014-04-01 Published:2013-12-09

RichHTML

1

PDF

678

Cited

6

Abstract: 【Objective】 The aim of this study was to clone a plasma membrane aquaporin gene EjPIP1 from Eriobotrya japonica variety ‘Zaozhong 6’, and to analyze its sequence characteristics, then to study the effects of AM fungi on the PIP expression pattern and water use efficiency(WUE). 【Method】 A pot experiment with seedlings of loquat cultivar (‘Zaozhong 6’), two treatments (AM and NM) and five replicates were conducted. The leaf water use efficiency was measured by using LI-6400 photosynthesis determination system. The full-length cDNA of EjPIP1 was obtained by the technology of reverse transcription PCR (RT-PCR) and RACE. The cDNA sequence and the deduced amino acid sequence were analyzed by bioinformatics method. The expression patterns of EjPIP1 in leaves and roots by AM fungi inoculation were determined by real-time RT-PCR.【Result】In the effective photosynthetic time(7:00—17:00), the water use efficiency of loquat leaves were significantly increased by AM fungi inoculation. Loquat plasma membrane aquaporin gene EjPIP1 (GenBank Accessin No. JX041627) was cloned. The full-length cDNA of EjPIP1 gene consists of 1 142 bp and contains a 861 bp open reading frame (ORF) encoding 286 amino acid proteins. Two highly conserved NPA (Asn-Pro-Ala, asparagine-proline-alanine) motifs of aquaporin were identified in EjPIP1. The homologue analysis revealed that the amino acids sequence of loquat PIP1 was highly homologous with other species, especially apple (Malus domestica), peach (Prunus persica) and strawberry (Fragaria chiloensis), which was up to 97%, 92% and 90%, respectively. Real-time PCR analysis revealed that EjPIP1 could be expressed in different loquat tissues including root and leaf, and the relative expression in leaf was higher than that in root. Furthermore, under well-watered condition, the expression of EjPIP1 in loquat leaf was significantly down-regulated by AM fungi inoculation, while the expression of EjPIP1 in root was up-regulated. 【Conclusion】 The water use efficiency of loquat leaves was significantly increased by AM fungi inoculation. A plasma membrane intrinsic proteins (PIPs) gene, designated as EjPIP1, was cloned from leaf of loquat (E. japonica ‘Zaozhong 6’). And the expression of this corresponding gene in loquat leaves and roots were influenced by AM fungi, which helped to improve water use efficiency of loquat seedlings after being inoculated with AM fungi.

Key words: Eriobotrya japonica , arbuscular mycorrhizal , aquaporin , cloning , expression analysis

[1]Maurel C, Verdoucq L, Luu D T, Santoni V. Plant aquaporins: membrane channels with multiple integrated functions. Annual Review of Plant Biology, 2008, 59:595-624.

[2]Heinen R B, Ye Q, Chaumont F. Role of aquaporins in leaf physiology. Journal of Experimental Botany, 2009, 60(11):2971-2985.

[3]庞永奇, 王高奇, 王旭初, 陈珈, 王学臣.植物水孔蛋白的亚细胞分布与生理功能研究浅析.生物化学与生物物理进展, 2012, 39(10):962-971.

Pang Y Q, Wang G Q, Wang X C, Chen J, Wang X C. A brief analysis of subcellular distribution and physiological functions of plant aquaporins. Progress in Biochemistry and Biophysics, 2012, 39(10):962-971. (in Chinese)

[4]Chaumont F, Barrieu F, Jung R, Chrispeels M J. Plasma membrane intrinsic proteins from maize cluster in two sequence subgroups with differential aquaporin activity. Plant Physiology, 2000, 122(4): 1025-1034.

[5]Chaumont F, Barrieu F, Wojcik E, Chrispeels M J, Jung R. Aquaporins constitute a large and highly divergent protein family in maize. Plant Physiology, 2001, 125:1206-1215.

[6]Johanson U, Karlsson M, Johansson I, Gustavsson S, Sjövall S, Fraysse L, Weig A R, Kjellbom P. The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a frame work for a new nomenclature for major intrinsic proteins in plants. Plant Physiology, 2001, 126:1358-1369.

[7]Sakurai J, Ishikawa F, Yamaguchi T, Uemura M, Maeshima M. Identification of 33 rice aquaporin genes and analysis of their expression and function. Plant and Cell Physiology, 2005, 46(9): 1568-1577.

[8]Augé R M. Water relations, drought and vesicular-arbuscular mycorrhizal symbiosis. Mycorrhiza, 2001, 11(1):3-42.

[9]Smith S E, Read D J. Mycorrhizal Symbiosis. London: Academic Press, 2008, 11-145.

[10]Ruiz-Lozano J M, Aroca R. Host response to osmotic stresses: stomatal behaviour and water use efficiency of arbuscular mycorrhizal plants//Koltai H, Kapulnik Y. eds. Arbuscular Mycorrhizas: Physiology and Function. Berlin: Springer-Verlag, 2010, 239-256.

[11]Yooyongwech S, Horigane A K, Yoshida M, Yamaguchi M, Sekozawa Y, Sugaya S, Gemma H. Changes in aquaporin gene expression and magnetic resonance imaging of water status in peach tree flower buds during dormancy. Physiologia Plantarum, 2008, 134(3):522-533.

[12]Picaud S, Becq F, Dédaldéchamp F, Ageorges A, Delrot S. Cloning and expression of two plasma membrane aquaporins expressed during the ripening of grape berry. Functional Plant Biology, 2003, 30(6):621-630.

[13]陈虎, 何新华, 罗聪, 邓立宝, 胡颖, 李明娟, 杨丽涛. 龙眼水通道蛋白基因 (DLPIP1) 的克隆与表达分析. 果树学报, 2012, 29(2):225-230.

Chen H, He X H, Luo C, Deng L B, Hu Y, Li M J, Yang L T. Cloning and expression of longan aquaporin (DLPIP1) gene. Journal of Fruit Science, 2012, 29(2):225-230. (in Chinese)

[14]Phillips J M, Hayman D S. Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Transactions of the British Mycological Society, 1970, 55(1):158-161.

[15]Giovannetti M, Mosse B. An evaluation of techniques for measuring vesicular arbuscular mycorrhizal infection in roots. New Phytologist, 1980, 84(3):489-500.

[16]Forrest K L, Bhave M. The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features. Functional & Integrative Genomics, 2008, 8(2):115-133.

[17]Fu X, Kong W, Peng G, Zhou J, Azam M, Xu C, Don G, Chen K. Plastid structure and carotenogenic gene expression in red-and white-fleshed loquat (Eriobotrya japonica) fruits. Journal of Experimental Botany, 2012, 63(1):341-354.

[18]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(4):402-408.

[19]Postaire O, Verdoucq L, Maurel C. Aquaporins in plants: from molecular structures to integrated functions. Advances in Botanical Research, 2008, 46:75-136.

[20]Wudick M M, Luu D T, Maurel C. A look inside: localization patterns and functions of intracellular plant aquaporins. New Phytologist, 2009, 184(2):289-302.

[21]林燕飞, 李红梅, 丁岳练, 黄新敏, 冼锡金, 何生根. 唐菖蒲质膜水孔蛋白基因GhPIP1;1的克隆及表达分析. 园艺学报, 2013, 40(1):145-154.

Lin Y F, Li H M, Ding Y L, Huang X M, Xian X J, He S G. Cloning and expression analysis of a plasma membrane aquaporin gene GhPIP1;1 in cut Gladiolus flowers. Acta Horticulturae Sinica, 2013, 40(1):145-154. (in Chinese)

[22]Comparot S, Morillon R, Badot P M. Water permeability and revolving movement in Phaseolus vulgaris L. twining shoots. Plant and Cell Physiology, 2000, 41(1):114-118.

[23]Alexandersson E, Fraysse L, Sjövall-Larsen S, Gustavsson S, Fellert M, Karlsson M, Johanson U, Kjellbom P. Whole gene family expression and drought stress regulation of aquaporins. Plant Molecular Biology, 2005, 59(3):469-484.

[24]Aharon R, Shahak Y, Wininger S, Bendov R, Kapulnik Y, Galili G. Overexpression of a plasma membrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but not under drought or salt stress. The Plant Cell, 2003, 15(2):439-447.

[25]Roussel H, Bruns S, Gianinazzi-Pearson V, Hahlbrock K, Franken P. Induction of a membrane intrinsic protein-encoding mRNA in arbuscular mycorrhiza and elicitor-stimulated cell suspension cultures of parsley. Plant Science, 1997, 126(2):203-210.

[26]Porcel R, Aroca R, Azcon R, Ruiz-Lozano J M. PIP aquaporin gene expression in arbuscular mycorrhizal Glycine max and Lactuca sativa plants in relation to drought stress tolerance. Plant Molecular Biology, 2006, 60(3):389-404.

[27]Aroca R, Porcel R, Ruiz-Lozano J M. How does arbuscular mycorrhizal symbiosis regulate root hydraulic properties and plasma membrane aquaporins in Phaseolus vulgaris under drought, cold or salinity stresses? New Phytologist, 2007, 173(4):808-816.

[28]李涛, 陈保冬. 丛枝菌根真菌通过上调根系及自身水孔蛋白基因表达提高玉米抗旱性. 植物生态学报, 2012, 36(9): 973-981.

Li T, Chen B D. Arbuscular mycorrhizal fungi improving drought tolerance of maize plants by up-regulation of aquaporin gene expressions in roots and the fungi themselves. Chinese Journal of Plant Ecology, 2012, 36(9): 973-981. (in Chinese)

[29]Zézé A, Brou Y C, Meddich A, Marty F. Molecular identification of MIP genes expressed in the roots of an arbuscular mycorrhizal Trifolium alexandrium L. under water stress. African Journal of Agricultural Research, 2008, 3(1), 78-83.

[30]Ruiz-Lozano J M, Alguacil M M, Bárzana G, Vernieri P, Aroca R. Exogenous ABA accentuates the differences in root hydraulic properties between mycorrhizal and non mycorrhizal maize plants through regulation of PIP aquaporins. Plant Molecular Biology, 2009, 70: 565-579.

[31]Bolandnazar S, Aliasgarzad N, Neishabury M R, Chaparzadeh N. 2007. Mycorrhizal colonization improves onion (Allium cepa L.) yield and water use efficiency under water deficit condition. Scientia Horticulturae, 2007, 114(1): 11-15.

[32]Kaya C, Higges D, Kirnak H, Tas I. Mycorrhizal colonization improves fruit yield and water use efficiency in watermelon (Citrullus Lanatus Thumb.) grown under well-watered and water-stress conditions. Plant and Soil, 2003, 253(2): 287-292.

[33]Birhane E, Sterck F J, Fetene M, Bongers F, Kuyper T W. Arbuscular mycorrhizal fungi enhance photosynthesis, water use efficiency, and growth of frankincense seedlings under pulsed water availability conditions. Oecologia, 2012, 169(4): 895-904.

[34]Sade N, Gebretsadik M, Seligmann R, Schwartz A, Wallach R, Moshelion M. The role of tobacco aquaporin1 in improving water use efficiency, hydraulic conductivity, and yield production under salt stress. Plant Physiology, 2010, 152(1):245-254.

[35]Krajinski F, Biela A, Schubert D, Gianinazzi-Pearson V, Kaldenhoff R, Franken P. Arbuscular mycorrhiza development regulates the mRNA abundance of Mtaqp 1 encoding a mercury-insensitive aquaporin of Medicago truncatula. Planta, 2000, 211(1):85-90.
[1] 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.
[2] 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.
[3] 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.
[4] 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.
[5] ZOU PeiYi, LIU MeiYan, WANG Ying, LI RanHong. Cloning and Functional Study of AkNAC2 from Actinidia kolomikta [J]. Scientia Agricultura Sinica, 2025, 58(19): 3985-3999.
[6] YI ZeHui, WANG Ying, SONG HuiXia, ZHAO Jing, MAO LiPing. Genome-Wide Identification and Expression Analysis of Peroxiredoxins Gene Family in Asparagus officinalis [J]. Scientia Agricultura Sinica, 2025, 58(18): 3728-3743.
[7] ZHANG ShuHong, GAO FengJu, WU QiuYing, JI JingXin, ZHANG YunFeng, XU Ke, GU ShouQin, FAN YongShan. Cloning and Expression Analysis of Heat Shock Protein HSP 9/12 Genes in Setosphaeria turcica [J]. Scientia Agricultura Sinica, 2025, 58(18): 3648-3663.
[8] QI XiangYu, LI XinRu, CHEN ShuangShuang, FENG Jing, CHEN HuiJie, LIU XinTong, JIN YuYan, DENG YanMing. Identification of the FLA Gene Family and Functional Analysis of JsFLA2 in Jasminum sambac [J]. Scientia Agricultura Sinica, 2025, 58(17): 3516-3530.
[9] ZENG YueHui, ZOU WenGuang, ZHAO FuMing, XIAO ChangChun, HUANG JianHong, MA BinLin, YANG WangXing, WEI XinYu, XU XuMing. Map-Based Cloning and Functional Verification of A Novel Split Glume Gene OsSG2 in Rice (Oryza sativa L.) [J]. Scientia Agricultura Sinica, 2025, 58(11): 2062-2080.
[10] DU JiaQi, ZHANG ZiWei, WANG RuoFei, LI Xing, GUO HongYan, YANG Shuo, FENG Cheng, HE TangQing, Giri Bhoopander, ZHANG XueLin. The Interactive Effects of Organic Fertilizer Substituting Chemical Fertilizers and Arbuscular Mycorrhizal Fungi on Soil Nitrous Oxide Emission in Shajiang Black Soil and Fluvo-Aquic Soil [J]. Scientia Agricultura Sinica, 2025, 58(1): 101-116.
[11] LÜ ShuWei, TANG Xuan, LI Chen. Research Progress on Seed Shattering of Rice [J]. Scientia Agricultura Sinica, 2025, 58(1): 1-9.
[12] WANG Wei, WU ChuanLei, HU XiaoYu, LI JiaJia, BAI PengYu, WANG GuoJi, MIAO Long, WANG XiaoBo. Genome-Wide Identification of Soybean LOX Gene Family and the Effect of GmLOX15A1 Gene Allele on 100-Seed Weight [J]. Scientia Agricultura Sinica, 2025, 58(1): 10-29.
[13] GUAN ZhiLin, JIN FengWei, LIU TingTing, WANG Yi, TAN YingYing, YANG ChunHui, LI RuiTong, WANG Bo, LIU KeDe, DONG Yun. Genetic Analysis and Gene Mapping of Glossy Leaf in Brassica napus [J]. Scientia Agricultura Sinica, 2024, 57(4): 650-662.
[14] TAN FangDai, HE YingXia, LIU JiaYue, LI AiHua, TAO YongSheng. Multidimensional Characterization of Astringency Quality in Dry Red Wine and Its Effects [J]. Scientia Agricultura Sinica, 2024, 57(21): 4342-4355.
[15] SUN SiSi, MA Wu, SI HuiRu, WANG XianZhong, LIU Qiang, LUO YanLin, CHEN XiaoYuLong, TANG Bin. AQPs Characteristics of Megoura crassicauda and Their Expression Changes in Response to High Relative Humidity Stress [J]. Scientia Agricultura Sinica, 2024, 57(20): 4057-4070.
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