甘蓝自交不亲和性相关基因BoGSTL21的克隆与表达分析

doi:10.3724/SP.J.1006.2020.04004

Abstract

Abstract:

Glutathione-S-transferase (GSTs) plays an important role in plant resistance to stress, cytotoxic release and plant growth and development. In this study, we identified an up-regulated gene named BoGSTL21 based on the stigma transcriptome data in 0-60 min self-pollination. BoGSTL21 had an open reading frame (ORF) with the length of 900 bp, encoded a protein of 299 amino acid residues, which contained GST-N and GST-C domains without signal peptide and transmembrane domain, the theory isoelectric point of BoGSTL21 was 8.49. The promoter of BoGSTL21 gene contained many cis-acting elements such as light response, auxin response, abscisic acid response, low temperature and drought response. BoGSTL21 expresses in different tissues of Brassica oleracea. The expression level in stigma varies with developmental time, and was mainly overexpressed in mature stigma. The results of qRT-PCR revealed that BoGSTL21 mRNA expression level after self-and cross-pollinations for 0 min to 60 min was consistent with that of RNA-seq data. It was found through yeast two-hybrid that BoGSTL21 protein interacted with pollen development-related protein BoFAB1C, auxin-related protein BoPATL2, and aldolase-type TIM barrel family protein BoF9N12_9. BoGSTL21 gene was successfully induced and expressed in E. coli BL21 (DE3) with a purified protein size of 34 kD, which was consistent with the predicted results. According to the above results, BoGSTL21 may be a novel protein involved in the SI response process, which provides a new content for further research and utilization of self-incompatibility in Brasscia oleracea.

Key words: Brassica oleracea, gene cloning, self-incompatibility, expression analysis, yeast two-hybrid

Tong-Hong ZUO, He-Cui ZHANG, Qian-Ying LIU, Xiao-Ping LIAN, Qin-Qin XIE, Deng-Ke HU, Yi-Zhong ZHANG, Yu-Kui WANG, Xiao-Jing BAI, Li-Quan ZHU. Molecular cloning and expression analysis of BoGSTL21 in self-incompatibility Brasscia oleracea[J].Acta Agronomica Sinica, 2020, 46(12): 1850-1861.

Figures/Tables 14

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Table 2

Fig. 6

Fig. 7

Fig. 8

Table 3

Table 4

Fig. 9

Fig. 10

References 41

[1]	Hamamura Y, Nagahara S, Higashiyama T . Double fertilization on the move. Curr Opin Plant Biol, 2012,15:70-77. doi: 10.1016/j.pbi.2011.11.001 pmid: 22153653
[2]	Gu T, Mazzurco M, Sulaman W, Matias D D, Goring D R . Binding of an arm repeat protein to the kinase domain of the S-locus receptor kinase. Proc Natl Acad Sci USA, 1998,95:382-387. doi: 10.1073/pnas.95.1.382 pmid: 9419384
[3]	Vanoosthuyse V, Tichtinsky G, Dumas C, Gaude T, Cock J M . Interaction of calmodulin, a sorting nexin and kinase-associated protein phosphatase with the Brassica oleracea S-locus receptor kinase. Plant Physiol, 2003,133:919-929. doi: 10.1104/pp.103.023846 pmid: 14555783
[4]	Stone S L, Anderson E M, Mullen R T, Goring D R . ARC1 is an E3 ubiquitin ligase and promotes the ubiquitination of proteins during the rejection of self-incompatible Brassica pollen. Plant Cell, 2003,15:885-898. doi: 10.1105/tpc.009845 pmid: 12671085
[5]	Nasrallah M E, Liu P, Nasrallah J B . Generation of self- incompatible Arabidopsis thaliana by transfer of two S-locus genes from A. lyrata. Science, 2002,297:247-249. doi: 10.1126/science.1072205 pmid: 12114625
[6]	Nasrallah M, Liu P, Sherman-Broyles S, Boggs N, Nasrallah J . Natural variation in expression of self-incompatibility in Arabidopsis thaliana: implications for the evolution of selfing. Proc Natl Acad Sci USA, 2004,101:16070-16074. doi: 10.1073/pnas.0406970101 pmid: 15505209
[7]	Nasrallah J B, Nasrallah M E . Robust self-incompatibility in the absence of a functional ARC1 gene in Arabidopsis thaliana. Plant Cell, 2014,26:3838-3841. doi: 10.1105/tpc.114.129387
[8]	Shimabukuro R H, Swanson H R, Walsh W C . Glutathione conjugation:atrazine detoxification mechanism in corn. Plant Physiol, 1970,46:103-107. doi: 10.1104/pp.46.1.103 pmid: 16657398
[9]	Dixon D P, Lapthorn A, Edwards R . Plant glutathione transferases. Methods Enzymol, 2005,401:169-186. doi: 10.1016/S0076-6879(05)01011-6 pmid: 16399386
[10]	Moons A . Osgstu3 and osgtu4,encoding tau class glutathione S-transferases, are heavy metal-and hypoxic stress-induced and differentially salt stress-responsive in rice roots. FEBS Lett, 2003,553:427-432. doi: 10.1016/s0014-5793(03)01077-9 pmid: 14572664
[11]	Soranzo N, Sari Gorla M, Mizzi L, De Toma G, Frova C . Organisation and structural evolution of the rice glutathione S-transferase gene family. Mol Genet Genomics, 2004,271:511-521. doi: 10.1007/s00438-004-1006-8 pmid: 15069639
[12]	Dixon D P, Davis B G, Edwards R . Functional divergence in the glutathione transferase superfamily in plants. J Biol Chem, 2002,277:30859-30869. doi: 10.1074/jbc.M202919200 pmid: 12077129
[13]	Moons A . Regulatory and functional interactions of plant growth regulators and plant glutathione S-transferases (GSTs). Vitam Horm, 2005,72:155-202. doi: 10.1016/S0083-6729(05)72005-7 pmid: 16492471
[14]	Marrs K . The functions and regulation of glutathione S-transferases in plants. Annu Rev Plant Physiol Plant Mol Biol, 1996,47:127-158. doi: 10.1146/annurev.arplant.47.1.127 pmid: 15012285
[15]	Edwards R, Dixon D P, Walbot V . Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci, 2000,5:193-198. doi: 10.1016/s1360-1385(00)01601-0 pmid: 10785664
[16]	Dixon D P, Cole D J, Edwards R . Dimerisation of maize glutathione transferases in recombinant bacteria. Plant Mol Biol, 1999,40:997-1008. doi: 10.1023/a:1006257305725 pmid: 10527424
[17]	Sommer A, Boger P . Characterization of recombinant corn glutathione S-transferase isoforms I, II, III, and IV. Pestic Biochem Physiol, 1999,63:127-138. doi: 10.1006/pest.1999.2396
[18]	胡廷章, 周大祥, 罗凯 . 植物谷胱甘肽转移酶的结构与功能及其基因表达. 植物生理学通讯, 2007,43:195-200.
	Hu T Z, Zhou D X, Luo K . Structure and biological function of glutathione transferases and their genes in plants. J Plant Physiol, 2007,43:195-200 (in Chinese with English abstract).
[19]	Sari-Gorla M, Ferrario S, Rossini L, Frova C, Villa M . Developmental expression of glutathione S-transferase in maize and its possible connection with herbicide tolerance. Euphytica, 1993,67:221-230. doi: 10.1007/BF00040624
[20]	Gietz R D, Schiestl R H . Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method. Nat Protocols, 2007,2:1-4. doi: 10.1038/nprot.2007.17 pmid: 17401330
[21]	Zeng J, Gao Q G, Shi S M, Lian X P, Converse R, Zhang H C, Yang X H, Ren X S, Chen S, Zhu L Q . Dissecting pistil responses to incompatible and compatible pollen in self-incompatibility Brassica oleracea using comparative proteomics. Protein J, 2017,36:123-137. doi: 10.1007/s10930-017-9697-y pmid: 28299594
[22]	Grotewold E . Subcellular trafficking of phytochemicals. Recent Res Dev Plant Physiol, 2001,2:31-48.
[23]	Alfenito M R, Souer E, Goodman C D, Buell R, Mol J, Koes R, Walbot V . Functional complementation of anthocyanin sequestration in the vacuole by widely divergent glutathione S-transferases. Plant Cell, 1998,10:1135-1149. doi: 10.1105/tpc.10.7.1135 pmid: 9668133
[24]	Mueller L A, Goodman C D, Silady R A, Walbot V . AN9, a petunia glutathione S-transferase required for anthocyanin sequestration, is a flavonoid-binding protein. Plant Physiol, 2000,123:1561-1570. doi: 10.1104/pp.123.4.1561 pmid: 10938372
[25]	Zhao J, Huhman D, Shadle G, He X Z, Sumner L W, Tang Y H, Dixon R A . MATE2 mediates vacuolar sequestration of flavonoid glycosides and glycoside malonates in Medicago truncatula. Plant Cell, 2011,23:1536-1555. doi: 10.1105/tpc.110.080804
[26]	Kitamura S, Shikazono N, Tanaka A . TRANSPARENT TESTA 19 is involved in the accumulation of both anthocyanins and proanthocyanidins in Arabidopsis. Plant J, 2004,37:104-114. doi: 10.1046/j.1365-313x.2003.01943.x pmid: 14675436
[27]	Hahlbrock K, Scheel D . Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol, 1989,40:347-369. doi: 10.1146/annurev.pp.40.060189.002023
[28]	Mo Y Y, Nagel C, Taylor L P . Biochemical complementation of chalcone synthase mutants defines a role for flavonols in functional pollen. Proc Natl Acad Sci USA, 1992,89:7213-7217. doi: 10.1073/pnas.89.15.7213 pmid: 11607312
[29]	Taylor L P, Grotewold E . Flavonoids as developmental regulators. Curr Opin Plant Biol, 2005,8:317-323. doi: 10.1016/j.pbi.2005.03.005 pmid: 15860429
[30]	Thieme C J, Rojas-Triana M, Stecyk E, Schudoma C, Zhang W, Yang L, Minambres M, Walther D, Schulze W X, Paz-Ares J, Scheible W R, Kragler F . Endogenous Arabidopsis messenger RNAs transported to distant tissues. Nat Plants, 2015,1:15025. doi: 10.1038/nplants.2015.25 pmid: 27247031
[31]	Hirano T, Matsuzawa T, Takegawa K, Sato M H . Loss-of- function and gain-of-function mutations in FAB1A/B impair endomembrane homeostasis, conferring pleiotropic developmental abnormalities in Arabidopsis. Plant Physiol, 2011,155:797-807. doi: 10.1104/pp.110.167981
[32]	Whitley P, Hinz S, Doughty H . Arabidopsis FAB1/PIKfyve proteins are essential for development of viable pollen. Plant Physiol, 2009,151:1812-1822. doi: 10.1104/pp.109.146159 pmid: 19846542
[33]	Tejos R, Rodriguez-Furlan C, Adamowski M, Sauer M, Norambuena L, Friml J . PATELLINS are regulators of auxin-mediated PIN1 relocation and plant development in Arabidopsis thaliana. J Cell Sci, 2018, 131: jcs204198. doi: 10.1242/jcs.204198 pmid: 28687624
[34]	Chen D, Zhao J . Free IAA in stigmas and styles during pollen germination and pollen tube growth of Nicotiana tabacum. Physiol Planta, 2008,134:202-215. doi: 10.1111/ppl.2008.134.issue-1
[35]	王玉奎, 张贺翠, 白晓璟, 廉小平, 施松梅, 刘倩莹, 左同鸿, 朱利泉 . 甘蓝BoPINs家族基因的特征和表达分析. 作物学报, 2019,45:1270-1278. doi: 10.3724/SP.J.1006.2019.84129
	Wang Y K, Zhang H C, Bai X J, Lian X P, Shi S M, Liu Q Y, Zuo T H, Zhu L Q . Characteristics and expression analysis of BoPINs family genes in Brassica oleracea. Acta Agron Sin, 2019,45:1270-1278 (in Chinese with English abstract).
[36]	Hasenstein K H, Zavada M S . Auxin modification of the incompatibility response in Theobroma cacao. Physiol Plant, 2001,112:113-118. doi: 10.1034/j.1399-3054.2001.1120115.x pmid: 11319022
[37]	Aloni R, Aloni E, Langhans M, Ullrich C I . Role of auxin in regulating Arabidopsis flower development. Planta, 2006,223:315-328. doi: 10.1007/s00425-005-0088-9
[38]	Tantikanjana T, Nasrallah J B . Non-cell-autonomous regulation of crucifer self-incompatibility by auxin response factor ARF3. Proc Natl Acad Sci USA, 2012,109:19468-19473. doi: 10.1073/pnas.1217343109 pmid: 23129621
[39]	Zettl R, Schell J, Palme K . Photo affinity labeling of Arabidopsis thaliana plasma membrane vesicles by 5-azido-[7- ³H] indole- 3-acetic acid: Identification of a glutathione S-transferase. Proc Natl Acad Sci USA, 1994,91:689-693. doi: 10.1073/pnas.91.2.689 pmid: 8290582
[40]	高世超, 林义章, 钟凤林, 赵瑞丽, 林琳琳, 占丽英 . 青花菜谷胱甘肽-S-转移酶基因克隆及其表达分析. 西北植物学报, 2014,34:651-657.
	Gao S C, Lin Y Z, Zhong F L, Zhao R L, Lin L L, Zhan L Y . Cloning of GST and its expression in Broccoli (Brassica oleracea var. italic). Acta Bot Boreali-Occident Sin, 2014,34:651-657 (in Chinese with English abstract).
[41]	Jiang J J, Jiang J X, Qiu L, Miao Y, Yao L N, Cao J S . Identification of gene expression profile during fertilization in Brassica campestris subsp. chinensis. Genome, 2012,56:39-48. doi: 10.1139/gen-2012-0088 pmid: 23379337

Related Articles 15

[1]	CUI Lian-Hua, ZHAN Wei-Min, YANG Lu-Hao, WANG Shao-Ci, MA Wen-Qi, JIANG Liang-Liang, ZHANG Yan-Pei, YANG Jian-Ping, YANG Qing-Hua. Molecular cloning of two maize (Zea mays) ZmCOP1 genes and their transcription abundances in response to different light treatments [J]. Acta Agronomica Sinica, 2022, 48(6): 1312-1324.
[2]	CHEN Song-Yu, DING Yi-Juan, SUN Jun-Ming, HUANG Deng-Wen, YANG Nan, DAI Yu-Han, WAN Hua-Fang, QIAN Wei. Genome-wide identification of BnCNGC and the gene expression analysis in Brassica napus challenged with Sclerotinia sclerotiorum and PEG-simulated drought [J]. Acta Agronomica Sinica, 2022, 48(6): 1357-1371.
[3]	ZHANG Yi-Zhong, ZENG Wen-Yi, DENG Lin-Qiong, ZHANG He-Cui, LIU Qian-Ying, ZUO Tong-Hong, XIE Qin-Qin, HU Deng-Ke, YUAN Chong-Mo, LIAN Xiao-Ping, ZHU Li-Quan. Codon usage bias analysis of S-locus genes SRK, SLG, and SP11/SCR in Brassica oleracea [J]. Acta Agronomica Sinica, 2022, 48(5): 1152-1168.
[4]	ZHOU Hui-Wen, QIU Li-Hang, HUANG Xing, LI Qiang, CHEN Rong-Fa, FAN Ye-Geng, LUO Han-Min, YAN Hai-Feng, WENG Meng-Ling, ZHOU Zhong-Feng, WU Jian-Ming. Cloning and functional analysis of ScGA20ox1 gibberellin oxidase gene in sugarcane [J]. Acta Agronomica Sinica, 2022, 48(4): 1017-1026.
[5]	JIN Min-Shan, QU Rui-Fang, LI Hong-Ying, HAN Yan-Qing, MA Fang-Fang, HAN Yuan-Huai, XING Guo-Fang. Identification of sugar transporter gene family SiSTPs in foxtail millet and its participation in stress response [J]. Acta Agronomica Sinica, 2022, 48(4): 825-839.
[6]	XIE Qin-Qin, ZUO Tong-Hong, HU Deng-Ke, LIU Qian-Ying, ZHANG Yi-Zhong, ZHANG He-Cui, ZENG Wen-Yi, YUAN Chong-Mo, ZHU Li-Quan. Molecular cloning and expression analysis of BoPUB9 in self-incompatibility Brassica oleracea [J]. Acta Agronomica Sinica, 2022, 48(1): 108-120.
[7]	YIN Ming, YANG Da-Wei, TANG Hui-Juan, PAN Gen, LI De-Fang, ZHAO Li-Ning, HUANG Si-Qi. Genome-wide identification of GRAS transcription factor and expression analysis in response to cadmium stresses in hemp (Cannabis sativa L.) [J]. Acta Agronomica Sinica, 2021, 47(6): 1054-1069.
[8]	ZUO Xiang-Jun, FANG Peng-Peng, LI Jia-Na, QIAN Wei, MEI Jia-Qin. Characterization of aphid-resistance of a hairy wild Brassica oleracea taxa, B. incana [J]. Acta Agronomica Sinica, 2021, 47(6): 1109-1113.
[9]	TANG Rui-Min, JIA Xiao-Yun, ZHU Wen-Jiao, YIN Jing-Ming, YANG Qing. Cloning of potato heat shock transcription factor StHsfA3 gene and its functional analysis in heat tolerance [J]. Acta Agronomica Sinica, 2021, 47(4): 672-683.
[10]	JIA Xiao-Ping, LI Jian-Feng, ZHANG Bo, QUAN Jian-Zhang, WANG Yong-Fang, ZHAO Yuan, ZHANG Xiao-Mei, WANG Zhen-Shan, SANG Lu-Man, DONG Zhi-Ping. Responsive features of SiPRR37 to photoperiod and temperature, abiotic stress and identification of its favourable allelic variations in foxtail millet (Setaria italica L.) [J]. Acta Agronomica Sinica, 2021, 47(4): 638-649.
[11]	YUE Jie-Ru, BAI Jian-Fang, ZHANG Feng-Ting, GUO Li-Ping, YUAN Shao-Hua, LI Yan-Mei, ZHANG Sheng-Quan, ZHAO Chang-Ping, ZHANG Li-Ping. Cloning and potential function analysis of ascorbic peroxidase gene of hybrid wheat in seed aging [J]. Acta Agronomica Sinica, 2021, 47(3): 405-415.
[12]	YANG Qin-Li, YANG Duo-Feng, DING Lin-Yun, ZHANG Ting, ZHANG Jun, MEI Huan, HUANG Chu-Jun, GAO Yang, YE Li, GAO Meng-Tao, YAN Sun-Yi, ZHANG Tian-Zhen, HU Yan. Identification of a cotton flower organ mutant 182-9 and cloning of candidate genes [J]. Acta Agronomica Sinica, 2021, 47(10): 1854-1862.
[13]	HE Xiao, LIU Xing, XIN Zheng-Qi, XIE Hai-Yan, XIN Yu-Feng, WU Neng-Biao. Molecular cloning, expression, and enzyme kinetic analysis of a phenylalanine ammonia-lyase gene in Pinellia ternate [J]. Acta Agronomica Sinica, 2021, 47(10): 1941-1952.
[14]	WANG Zhen, YAO Meng-Nan, ZHANG Xiao-Li, QU Cun-Min, LU Kun, LI Jia-Na, LIANG Ying. Prokaryotic expression, subcellular localization and yeast two-hybrid library screening of BnMAPK1 in B. napus [J]. Acta Agronomica Sinica, 2020, 46(9): 1312-1321.
[15]	LI Guo-Ji, ZHU Lin, CAO Jin-Shan, WANG You-Ning. Cloning and functional analysis of GmNRT1.2a and GmNRT1.2b in soybean [J]. Acta Agronomica Sinica, 2020, 46(7): 1025-1032.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

引物名称 Primer name	引物序列 Primer sequence (5°-3°)	用途 Functions
BoGSTL21-PGEX-4T-1	F: GATCTGGTTCCGCGTGGATCCATGAGTGCCGGAGTGAGAGTTAG	基因的原核表达
	R: CTCGAGTCGACCCGGGAATTCGGGACGTGCTTCTGCTTGG	Prokaryotic expression
qRT-PCR	F: TTCCTTTGCCGATTTAGTTTGG	荧光定量PCR引物
	R: AGTGTTCATCTCCTTAAGCCAA	qRT-PCR
dActin	F: GGCTGATGGTGAAGATATTCA	内参引物
	R: CAAGCACAATACCAGTAGTAC	Internal reference primers
BoGSTL21-BK	F: TCAGAGGAGGACCTGCATATGAGTGCCGGAGTGAGAGTTA	酵母双杂交引物 Yeast two-hybrid primers
	R: TCGACGGATCCCCGGGAATTCGGGACGTGCTTCTGCTTG
BoFAB1C-AD	F: GTACCAGATTACGCTCATATGGGGATGGTGAAGTTCTCTGTG
	R: ATGCCCACCCGGGTGGAATTCGTTCCATGGCTCAGGAACC
BoPATL2-AD	F: GTACCAGATTACGCTCATATGGCTCAAGAAGAGATACAGAAG
	R: ATGCCCACCCGGGTGGAATTCTTATGCTTGCGTTTTGAACC
BoF9N12_9-AD	F: GTACCAGATTACGCTCATATGGTGGTGTCGCCAAAGATAG
	R: ATGCCCACCCGGGTGGAATTCTCAGGTGATGGGTTGGGC
1391	F: GAACTGATCGTTAAAACTGC	通用引物
	R: TGGTCTTCTGAGACTGTATC	Universal primer
BoGSTL21-GUS	F: CAAGCTTGGCTGCAGGTCGACATGTTATACGTTGCGAACGC	基因的启动子活性分析
	R: GGTGGACTCCTCTTAGAATTCACTCAATCGTTCTTCTTCCGT	Promoter activity analysis

相关功能预测 Associated putative function	启动子顺式作用元件 Cis-elements in the promoter region
脱落酸(ABA)应答 Abscisic acid response	ABRE
光响应 Light response	G-Box, GT1-motif, TCT-motif
低氧诱导 Anaerobic induction response	ARE
促进和增强基因转录 Promote and enhance gene transcription	CAAT-box
赤霉素应答 Gibberellin-response	GARE-motif, P-box
MYBHv1结合位点 MYBHv1 binding site	CCAAT-box
涉及光响应的MYBHv1结合位点 MYB binding site involved in light response	MRE
转录启动约-30的核心启动子 Core promoter element around -30 of transcription start	TATA-box
低温响应 Low-temperature response	LTR
干旱诱导 Drought induced response	MBS
水杨酸(SA)反应 Salicylic acid response	TCA-element
MeJA茉莉酸响应 MeJA-response	CGTCA-motif, TGACG-motif
生长素(IAA)应答 Auxin-response	TGA-element

候选蛋白Candidate protein	蛋白名称 Protein name	功能注释 Functional annotations
BoFAB1C	1-磷脂酰肌醇-3-磷酸5-激酶 1-phosphatidylinositol-3- phosphate 5-kinase	具有1-磷脂酰肌醇-3-磷酸5-激酶活性, ATP结合,参与磷脂酰肌醇磷酸化, 花粉发育, 气孔关闭的生物过程。 It has 1-phosphatidylinositol-3-phosphate 5-kinase activity, ATP binding, participates in the biological processes of phosphatidylinositol phosphorylation, pollen development, and stomatal closure.
BoPATL2	patellin-2	PATL属于具有高尔基动力学(GOLD)结构域和Sec14p-like结构域串联的蛋白质家族。PATL受生长素调节。 PATL belongs to a family of proteins with a Golgi dynamics (GOLD) domain and a Sec14p-like domain tandem. PATL is regulated by auxin.
BoF9N12_9	磷酸核糖3-差向异构酶 Ribulose-phosphate 3-epimerase	属于醛缩酶型TIM桶家族蛋白, 参与碳水化合物代谢过程, 磷酸戊糖途径, 氨基酸的生物合成。 Belongs to the aldolase type TIM barrel family protein, involved in carbohydrate metabolism, pentose phosphate pathway, amino acid biosynthesis.

编号 No.	酵母菌种(质粒) Yeast strain (plasmid)	培养基 Yeast medium	菌斑 Colony	颜色 Color
1	Y2HGold (pGADT7-T×pGBKT7-53)	SD/-Leu/-Trp	是Yes	白色White
2	Y2HGold (pGADT7-T×pGBKT7-Lam)	SD/-Leu/-Trp	是Yes	红色Red
3	Y2HGold (pGADT7-BoFAB1C×pGBKT7-BoGSTL21)	SD/-Leu/-Trp	是Yes	白色White
4	Y2HGold (pGADT7-BoPATL2×pGBKT7-BoGSTL21)	SD/-Leu/-Trp	是Yes	白色White
5	Y2HGold (pGADT7-BoF9N12×pGBKT7-BoGSTL21)	SD/-Leu/-Trp	是Yes	白色White
6	Y2HGold (pGADT7-T×pGBKT7-53)	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White
7	Y2HGold (pGADT7-T×pGBKT7-Lam)	SD/-Ade/-His/-Leu/-Trp	是Yes	无色No
8	Y2HGold (pGADT7-BoFAB1C×pGBKT7-BoGSTL21)	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White
9	Y2HGold (pGADT7-BoPATL2×pGBKT7-BoGSTL21)	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White
10	Y2HGold (pGADT7-BoF9N12×pGBKT7-BoGSTL21)	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White

Molecular cloning and expression analysis of BoGSTL21 in self-incompatibility Brasscia oleracea

RichHTML375

PDF