秀珍菇原基形成相关基因PpFBD1的克隆与表达研究

doi:10.3969/j.issn.1004-1524.2020.01.12

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (1): 93-97.DOI: 10.3969/j.issn.1004-1524.2020.01.12

秀珍菇原基形成相关基因PpFBD1的克隆与表达研究

王伟科¹, 宋吉玲¹, 陆娜¹, 袁卫东¹, 闫静¹, 陈观平²

1. 杭州市农业科学研究院,浙江杭州 310024;
2. 浙江省中医药研究院,浙江杭州 310012

收稿日期:2019-06-13 出版日期:2020-01-25 发布日期:2020-03-11
作者简介:王伟科(1981—),男,浙江宁海人,学士,高级农艺师,主要从事食用菌育种与栽培技术研究。E-mail:akeok@126.com
基金资助:
浙江省农业(食用菌)新品种选育重大科技专项(2016C02057); 浙江省科技计划(2016F10025)

Cloning and expression of the PpFBD1 involved in primordium formation of Pleurotus pulmonarius

WANG Weike¹, SONG Jiling¹, LU Na¹, YUAN Weidong¹, YAN Jing¹, CHEN Guanping²

1. Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China;
2. Zhejiang Academy of Chinese Medicine, Hangzhou 310012,China

Received:2019-06-13 Online:2020-01-25 Published:2020-03-11

摘要/Abstract

摘要： 前期研究中通过对秀珍菇经低温诱导后不同发育阶段样本进行转录组测序及差异表达基因分析发现,ID为Cluster-6377.64510的基因(命名为PpFBD1)在原基形成前期的样本中高表达。为进一步了解分析其表达特性及功能,根据转录组测序结果设计特异性引物,利用RT-PCR技术从秀珍菇中克隆获得了该基因的cDNA全长。该基因由342个核苷酸组成,编码一个由113个氨基酸组成的蛋白,蛋白相对分子质量约11 ku。进化树分析显示,PpFBD1编码蛋白与糙皮侧耳hydrophobin1编码的疏水蛋白亲缘关系最近。Gene Ontology功能分析表明,PpFBD1主要参与真菌类细胞壁的合成。荧光定量RT-PCR检测显示,该基因在菌丝体经低温诱导后恢复至室温阶段(原基形成前期)表达量最高,这表明其可能参与调控秀珍菇原基形成过程。本研究结果为阐明秀珍菇原基形成机制提供参考。

关键词: 秀珍菇, PpFBD1, 基因克隆, 序列分析, 表达分析

Abstract: Through transcriptome sequencing and differential expression gene analysis of Pleurotus pulmonarius samples at different developmental stages induced by low temperature in the previous study, it was found that the gene with ID Cluster-6377.64510 (named as PpFBD1) was highly expressed at the prophase of primordium formation. To further understand and analyze its expression characteristics and functions, the full-length cDNA sequences of PpFBD1 was cloned from Pleurotus pulmonarius by RT-PCR method with the specific primers designed based on transcriptome data. The PpFBD1 was composed of 342 nucleotides that encode a protein consisted of 113 amino acids. The relative molecular weight of this protein is about 11 ku. Phylogenetic tree analysis showed that the protein encoded by PpFBD1 had the closest relationship with hydrophobin1 of Pleurotus ostreatus. GO functional analysis showed that the PpFBD1 mainly involved in the synthesis of fungal cell walls. Fluorescence quantitative RT-PCR detection found that the expression of the gene was the highest in the phase of mycelium recovery to room temperature after induction at low temperature. It suggested that the gene may be involved in the regulation of primordium morphogenesis in Pleurotus pulmonarius. The results of this study provided a reference for elucidating the formation mechanism of the primordium of Pleurotus pulmonarius.

Key words: Pleurotus pulmonarius, PpFBD1, gene cloning, sequence analysis, expression analysis

中图分类号:

S646.9

王伟科, 宋吉玲, 陆娜, 袁卫东, 闫静, 陈观平. 秀珍菇原基形成相关基因PpFBD1的克隆与表达研究[J]. 浙江农业学报, 2020, 32(1): 93-97.

WANG Weike, SONG Jiling, LU Na, YUAN Weidong, YAN Jing, CHEN Guanping. Cloning and expression of the PpFBD1 involved in primordium formation of Pleurotus pulmonarius[J]. , 2020, 32(1): 93-97.

参考文献

[1] 宋林丽, 邢晓科, 郭顺星. 大型真菌子实体发生的形态学过程及调控机制[J]. 菌物学报, 2018, 37(6): 671-684.
SONG L L, XING X K, GUO S X.Morphological process and regulation mechanisms of fruiting body differentiation in macrofungi: a review[J]. Mycosystema, 2018, 37(6): 671-684.(in Chinese with English abstract)
[2] 马爱民, 关海山. 平菇子实体分化发育相关基因片段的克隆[J]. 华中农业大学学报, 2001, 20(4): 299-302.
MA A M, GUAN H S.Cloning of differentiation and development related gene fragments in Pleurotus ostreatus[J]. Journal of Huazhong Agricultural, 2001, 20(4): 299-302.(in Chinese with English abstract)
[3] YAMADA M, SAKURABA S, SHIBATA K, et al.Cloning and characterization of a gene coding for a hydrophobin, Fv-hyd1, specifically expressed during fruiting body development in the basidiomycete Flammulina velutipes[J]. Applied Microbiology and Biotechnology, 2005, 67(2): 240-246.
[4] 韩星, 魏仲阳, 张晔琳, 等. 金针菇寡肽转运蛋白编码基因fvopt1与fvopt2的鉴定与表达分析[J]. 食用菌学报, 2018, 25(4): 17-23.
HAN X, WEI Z Y, ZHANG Y L, et al.Identification and expression analysis of two oligopeptide transporter encoding genes fvopt1 and fvopt2 from Flammulina velutipes[J]. Acta Edulis Fungi, 2018, 25(4): 17-23.(in Chinese with English abstract)
[5] 仝宗军, 严俊杰, 张磊, 等. 金针菇转运蛋白基因fv-mfs1的序列与表达分析[J]. 食用菌学报, 2017, 24(3): 1-6.
TONG Z J, YAN J J, ZHANG L, et al.Sequence and expression analysis of transporter protein gene fv-mfs1 in Flammulina velutipes[J]. Acta Edulis Fungi, 2017, 24(3): 1-6.(in Chinese with English abstract)
[6] 周烁红, 沈颖越, 蔡为明, 等. 肺形侧耳变温结实相关基因Ppcsl-1的克隆及功能预测[J]. 菌物学报, 2016, 35(8): 946-955.
ZHOU S H, SHEN Y Y, CAI W M, et al.Cloning and functional prediction of the Ppcsl-1 related to change-temperature fruiting of Pleurotus pulmonarius[J]. Mycosystema, 2016, 35(8): 946-955.(in Chinese with English abstract)
[7] 闫述模, 袁媛, 杨滨, 等. 一种适用于不同发育期山楂果实总RNA提取的方法[J]. 中国实验方剂学杂志, 2016, 22(14): 39-43.
YAN S M, YUAN Y, YANG B, et al.Good method for isolating total RNA from Crataegi fructus at different developmental stages[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2016, 22(14): 39-43.(in Chinese with English abstract)
[8] 崔波, 王洁琼, 宋彩霞, 等. 蝴蝶兰SOC1基因的克隆及表达分析[J]. 分子植物育种, 2016, 14(3): 548-553.
CUI B, WANG J Q, SONG C X, et al.Cloning and expression analysis of SOC1 gene from Phalaenopsis[J]. Molecular Plant Breeding, 2016, 14(3): 548-553.(in Chinese with English abstract)
[9] 沈颖越, 顾敏, 蔡为明, 等. 秀珍菇子实体发育相关消减杂交cDNA文库的构建与分析[J]. 浙江农业学报, 2014, 26(2): 330-334.
SHEN Y Y, GU M, CAI W M, et al.Construction and analysis of a suppressive subtractive hybridisation cDNA library related to fruiting body formation of Pleurotus pulmonarius after cold stimulation[J]. Acta Agriculturae Zhejiangensis, 2014, 26(2): 330-334.(in Chinese with English abstract)
[10] SAKAMOTO Y, NAKADE K, SATO T.Characterization of the post-harvest changes in gene transcription in the gill of the Lentinula edodes fruiting body[J]. Current Genetics, 2009, 55(4): 409-423.
[11] 王伟科, 宋吉玲, 闫静, 等. 秀珍菇转录组测序和初步分析[J]. 南京农业大学学报, 2019, 42(2): 292-299.
WANG W K, SONG J L, YAN J, et al.Transcriptome sequencing and analysis of Pleurotus pulmonarius[J]. Journal of Nanjing Agricultural University, 2019, 42(2): 292-299.(in Chinese with English abstract)
[12] MORRIS V K, KWAN A H, MACKAY J P, et al.Backbone and sidechain¹H,¹³C and¹⁵N chemical shift assignments of the hydrophobin DewA from Aspergillus nidulans[J]. Biomolecular NMR Assignments, 2012, 6(1): 83-86.
[13] SUNDE M, KWAN A H Y, TEMPLETON M D, et al. Structural analysis of hydrophobins[J]. Micron, 2008, 39(7): 773-784.
[14] OHM R A, DE JONG J F, LUGONES L G, et al. Genome sequence of the model mushroom Schizophyllum commune[J]. Nature Biotechnology, 2010, 28(9): 957-963.
[15] 陈仁良, 李肖, 龙莹, 等. 金针菇疏水蛋白Fv-Hyd1及其潜在的转录因子Fv-Rtg3的共表达分析[J]. 浙江大学学报(农业与生命科学版), 2016, 42(3): 313-320.
CHEN R L, LI X, LONG Y, et al.Analysis on the co-expression of hydrophobin fv-Hyd1 and its potential transcription factor fv-Rtg3 in Flammulina velutipes[J]. Journal of Zhejiang University(Agriculture and Life Sciences), 2016, 42(3): 313-320.(in Chinese with English abstract)

秀珍菇原基形成相关基因PpFBD1的克隆与表达研究

Cloning and expression of the PpFBD1 involved in primordium formation of Pleurotus pulmonarius

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	何佳琦, 翟莹, 张军, 邱爽, 李铭杨, 赵艳, 张梅娟, 马天意. 大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达[J]. 浙江农业学报, 2021, 33(1): 1-7.
[2]	王伟科, 陆娜, 闫静, 宋吉玲, 袁卫东, 周祖法. 秀珍菇S-腺苷甲硫氨酸合成酶基因(PpSAMS)的克隆与表达分析[J]. 浙江农业学报, 2021, 33(1): 62-68.
[3]	刘凯, 冯晓宇, 马恒甲, 谢楠. 钱塘江三角鲂线粒体基因组测序及其结构特征分析[J]. 浙江农业学报, 2020, 32(9): 1591-1608.
[4]	倪征, 陈柳, 华炯钢, 叶伟成, 云涛, 朱寅初, 张存. 浙江省地方鸡种禽白血病毒抗原检测与部分分离株GP85基因序列分析[J]. 浙江农业学报, 2020, 32(8): 1357-1362.
[5]	袁献宇, 杨龙斌, 何赞赞, 毛天骄, 何长生, 占松鹤, 孙裴, 魏建忠, 李郁. 安徽省猪伪狂犬病毒的分离鉴定及其主要毒力基因分子特征[J]. 浙江农业学报, 2020, 32(1): 43-56.
[6]	德西措姆, 王印, 杨泽晓, 姚学萍, 罗燕, 廖倡宇, 张鹏飞, 江地科, 项明源, 姜瑞姣, 宋勇. 2016—2018年四川地区猪流行性腹泻病毒流行病学调查与ORF3、S1部分基因序列分析[J]. 浙江农业学报, 2019, 31(9): 1423-1428.
[7]	易可可, 阴文奇, 周远成, 蒋金蓁, 张白玉, 李中银, 颜其贵. 四株猪伪狂犬病毒株的分离鉴定与主要毒力基因分析[J]. 浙江农业学报, 2019, 31(9): 1429-1436.
[8]	刘加林, 刘士力, 蒋文枰, 程顺, 迟美丽, 郑建波, 贾永义, 赵金良, 尹绍武, 顾志敏. 河川沙塘鳢GH基因及侧翼的克隆与生物信息学分析[J]. 浙江农业学报, 2019, 31(9): 1461-1470.
[9]	王元红, 鲁智敏, 张学琪, 刘自敏, 胡子慧, 杨侃侃, 刘红梅, 潘玲, 彭开松, 李永东, 王勇. 一株J亚型禽白血病病毒的分离鉴定及其gp85基因的序列分析[J]. 浙江农业学报, 2019, 31(5): 709-715.
[10]	程亮. 青海省部分地区马铃薯Y病毒cp基因序列分析[J]. 浙江农业学报, 2019, 31(11): 1888-1895.
[11]	周会明, 张焱珍, 柴红梅, 杨荣情, 谭莹, 张萍萍, 白玉英, 赵一莲, 金玉洁. 一株临沧野生黄鸡?的鉴定与生长条件研究[J]. 浙江农业学报, 2019, 31(10): 1655-1662.
[12]	许青松, 赵佳, 魏运民, 韩蓉蓉, 刘卢生, 蒋曹德, 玉永雄. 紫花苜蓿MsOXO基因的克隆及表达分析[J]. 浙江农业学报, 2019, 31(1): 11-19.
[13]	王玉书, 王欢, 郭宇, 周明慧, 陈璐, 陈阳. 羽衣甘蓝β-胡萝卜素羟化酶基因的克隆及表达分析[J]. 浙江农业学报, 2019, 31(1): 80-85.
[14]	杨舟, 吕可, 吕珊, 王俊杰, 张荻. 百子莲2个ARF基因与2个Aux/IAA基因的全长克隆与序列分析[J]. 浙江农业学报, 2019, 31(1): 86-97.
[15]	巴少波, 石林, 李群景, 刘正奎, 陈琳, 王晓杜, 宋厚辉. 猪繁殖与呼吸综合征病毒17-ZJ-HZ毒株的分离鉴定与分子流行病学分析[J]. 浙江农业学报, 2018, 30(8): 1303-1311.