水稻干尖线虫Ab-lea基因在高渗透压下的表达
Expression Under Hypertonic Osmotic Stress of Ab-lea from Aphelenchoides besseyi
Online published: 2017-11-10
【目的】 水稻干尖线虫(Aphelenchoides besseyi)在侵染寄主以前,能够以变渗隐生状态在土壤和种子中的高渗透压环境下长期存活,使防治该线虫极为困难。通常,非生物胁迫下,胚胎发育晚期丰富蛋白(late embryogenesis abundant protein,LEA)的基因表达量会显著上调。作为逆境表达蛋白,LEA能在逆境下保护生物。为探究水稻干尖线虫LEA编码基因(Ab-lea)在高渗透压胁迫下的表达模式及功能,【方法】根据转录组数据克隆到Ab-lea。以无机溶剂饱和MgSO4·7H2O溶液为高渗透压胁迫剂,对水稻干尖线虫进行高渗透压胁迫,240 min后该线虫进入变渗隐生。通过RT-PCR验证Ab-lea在线虫进入变渗隐生及维持变渗隐生过程中的表达特性。使用RNAi技术,探究该线虫Ab-lea沉默后,高渗透压胁迫下存活率变化。【结果】 RT-PCR结果表明,Ab-lea在进入变渗隐生前期及维持变渗隐生过程中表达量上调。Ab-lea沉默后,水稻干尖线虫在进入变渗隐生后存活率显著下降。【结论】 表明该基因可能参与调控水稻干尖线虫变渗隐生。本研究为进一步探究该线虫抗高渗透压胁迫的生理机制奠定基础,为防治该线虫提供新思路。
关键词： 胚胎发育晚期丰富蛋白; 水稻干尖线虫; 高渗透压胁迫; 变渗隐生; 基因干涉
陈俏丽, 王峰, 李丹蕾, 零雅茗, 张瑞芝 . 水稻干尖线虫Ab-lea基因在高渗透压下的表达[J]. 中国水稻科学, 2017 , 31(6) : 652 -657 . DOI: 10.16819/j.1001-7216.2017.7004
【Objective】Rice white tip nematodes (Aphelenchoides besseyi) can enter osmobiosis to survive hypertonic osmotic stress in soil and seeds before infecting hosts, making the prevention and control of the nematode rather difficult. Normally, under hypertonic osmotic stress, the gene expression of late embryogenesis abundant protein (LEA) will significantly increase. As an adversity expressive protein, LEA can protect the organism under adversity. In order to study the expression pattern and the function of LEA gene from A. besseyi (Ab-lea) under hypertonic osmotic stress, 【Method】the gene was cloned according to the results of the transcriptome sequencing. A saturated MgSO4·7H2O solution was used as a hypertonic osmotic solution to dehydrate nematodes. The expression of Ab-lea was tested by RT-PCR during Osmobiosis. RNAi was used to investigate the survival of nematodes under hypertonic osmotic stress after silencing of Ablea.【Result】Nematodes will turn into osmobiosis after soaking in a saturated MgSO4·7H2O solution for 240 min. RT-PCR results showed that the expression of Ab-lea was upregulated when nematodes were at the beginning of hypertonic osmotic stress treatment and during the osmobiosis. 【Conclusion】The survival rate of the nematodes dropped significantly under osmobiosis after Ab-lea silence, which showed that Ab-lea may be involved in the regulation of osmobiosis of the nematode. It lays a foundation of further exploration for the physiological mechanism of nematode-resistance to hypertonic osmotic stress and provides new ideas for the prevention and control of the nematode.
| [1] | Fortuner R, Williams K J O. Review of the literature onAphelenchoides besseyi Christie, 1942, the nematode causing “White tip” disease in rice. Helminthol Abstr Ser B, Plant Nemat, 1975, 44(1): 1-40. |
| [2] | 裴艳艳,程曦,徐春玲,杨再福,谢辉. 中国水稻干尖线虫部分群体对水稻的致病力测定. 中国水稻科学,2012,26(2):218-226. |
| [2] | Pei Y Y, Cheng X, Xu C L, Yang Z F, Xie H.Virulence of part populations of Aphelenchoides besseyi on rice in China. Chin J Rice Sci, 2012, 26(2): 218-226.(in Chinese with English abstract) |
| [3] | Xun Y, Patrick D C, Richou H, John J, Shulong C, Maurice M.Osmotic responses of different strains of Steinernema carpocapsae. Nematol, 2011, 13(7): 845-851. |
| [4] | Glazer I, Salame L.Osmotic survival of the entomopathogenic nematodeSteinernema carpocapsae. Biol Control, 2000, 18(3): 251-257. |
| [5] | Dure L 3rd, Greenway S C, Galau G A. Developmental biochemistry of cottonseed embryogenesis and germination: Changing messenger ribonucleic acid populations as shown by in vitro and in vivo protein synthesis.Biochemistry, 1981, 20(14): 4162-4168. |
| [6] | Battaglia M, Covarrubias A A.The enigmatic LEA proteins and other hydrophilins.Plant Physiol, 2008, 148(1): 6-24. |
| [7] | Gal T Z, Glazer I, Koltai H.An LEA group 3 family member is involved in survival ofC. elegans during exposure to stress. FEBS Let, 2004, 577(1/2): 21-26. |
| [8] | Browne J A, Dolan K M, Tyson T, Goyal K, Tunnacliff A, Burnell A M.Dehydration-specific induction of hydrophilic protein genes in the anhydrobiotic nematodeAphelenchus avenae. Eukar Cell, 2004, 3(4): 966-975. |
| [9] | Adhikari B N, Wall D H, Adams B J.Desiccation survival in an Antarctic nematode: Molecular analysis using expressed sequenced tags.BMC Genom, 2009, 10(1): 1-18. |
| [10] | Tunnacliffe A, Lapinski J, Mcgee B.A putative LEA protein, but no trehalose, is present in anhydrobiotic bdelloid rotifers.Hydrobiol, 2005, 546(1): 315-321. |
| [11] | Kikawada T, Nakahara Y, Kanamori Y, Iwata K, Watanabe M, McGee B, Tunnacliff A, Okuda T. Dehydration-induced expression of LEA proteins in an anhydrobiotic chironomid.Biochem Biophysic Res Comm, 2006, 348(1): 56-61. |
| [12] | Hand S C, Jones D, Menze M A, Witt T L.Life without water: Expression of plant LEA genes by an anhydrobiotic arthropod.J Exp Zool Part A Ecol Genet Physiol, 2007, 307A(1): 62-66. |
| [13] | Liu G, Xu H, Zhang L, Zheng Y.Fe binding properties of two soybean (Glycine max L.) LEA4 proteins associated with antioxidant activity. Plant Cell Physio, 2011, 52(6): 994-1002. |
| [14] | Krüger C, Berkowitz O, Stephan U W, Hell R.A metal-binding member of the late embryogenesis abundant protein family transports iron in the phloem ofRicinus communis L. J Biol Chem, 2002, 277(28): 25062-9. |
| [15] | Goyal K, Walton L, Tunnacliffe A.LEA proteins prevent protein aggregation due to water stress.Biochem J, 2005, 388(388): 151-7. |
| [16] | Tunnacliffe A, Hincha D K, Leprince O, Macherel D.LEA proteins: versatility of form and function// Dormancy and Resistance in Harsh Environments. Springer: Berlin Heidelberg, 2010: 91-108. |
| [17] | Dure L 3rd, Crouch M, Harada J, Ho T H, Mundy J, Quatrano R, Thomas T, Sung Z R. Common amino acid sequence domains among the LEA proteins of higher plants.Plant Mol Bio, 1989, 12(5): 475-486. |
| [18] | Shih M D, Hoekstra F A, Hsing Y I C. Late embryogenesis abundant proteins.Adv Bot Res, 2008, 48(2008): 211-255. |
| [19] | Solomon A, Salomon R, Paperna I, Glazer I.Desiccation stress of entomopathogenic nematodes induces the accumulation of a novel heat-stable protein.Parasitology, 2000, 121(4): 409-416. |
| [20] | Dure Iii L.Occurrence of a repeating 11-mer amino acid sequence motif in diverse organisms.Prot Peptide Let, 2001, 8(2): 115-122. |
| [21] | Browne J, Tunnacliffe A, Burnell A.Anhydrobiosis: plant desiccation gene found in a nematode.Nature, 2002, 416(6876): 38. |
| [22] | Curry J, Walkersimmons M K.Unusual sequence of group 3 LEA (II) mRNA inducible by dehydration stress in wheat.Plant Mol Biol, 1993, 21(5): 907-912. |
| [23] | Hsing Y C, Chen Z Y, Shih M D, Hsieh J S, Chow T Y.Unusual sequences of group 3 LEA mRNA inducible by maturation or drying in soybean seeds.Plant Mol Biol, 1995, 29(4): 863-868. |
| [24] | Wang F, Li D, Wang Z, Dong A, Liu L, Wang B, Chen Q, Liu X.Transcriptomic analysis of the rice white tip nematode,Aphelenchoides besseyi(Nematoda: Aphelenchoididae). Plos One, 2014, 9(3): 131-134. |
| [25] | Wang F, Wang Z, Li D, Chen Q. Identification and characterization of a Bursaphelenchus xylophilus(Aphenlenchida: Aphelenchoididae) thermotolerance- related gene: Bx-HSP90. Inter J Mol Sci, 2012, 13: 8819-8833. |
| [26] | Perry R N. Desiccation survival of parasitic nematodes. Parasitology, 1999, 119(S1(Parasite Adaptation to Environmental Constraints)): 19-30. |
| [27] | Clegg J S.Cryptobiosis: a peculiar state of biological organization.Compar Biochem Physiol Part B Biochem Mole Biol, 2001, 128(4): 613-624. |
| [28] | Crowe J H, Hoekstra F A, Crowe L M.Anhydrobiosis. (1992).Ann Rev Physio, 1992, 54: 579-599. |
| [29] | Reardon W, Chakrabortee S, Pereira T C, Tyson T, Banton M C, Dolan K M, Culleton B A, Wise M J, Burnell A M, Tunnacliffe A.Expression profiling and cross-species RNA interference (RNAi) of desiccation-induced transcripts in the anhydrobiotic nematodeAphelenchus avenae. BMC Mol Biol, 2010, 11(1): 6. |
/
| 〈 |
|
〉 |