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作物学报

作物学报 ›› 2020, Vol. 46 ›› Issue (9): 1312-1321.doi: 10.3724/SP.J.1006.2020.04019

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜BnMAPK1的原核表达、亚细胞定位及酵母双杂交文库筛选

王珍(), 姚梦楠(), 张晓莉, 曲存民, 卢坤, 李加纳, 梁颖*()   

  1. 西南大学农学与生物科技学院 / 油菜工程研究中心 / 西南大学现代农业科学研究院, 重庆 400715
  • 收稿日期:2020-01-26 接受日期:2020-04-15 出版日期:2020-09-12 网络出版日期:2020-05-07
  • 通讯作者: 梁颖
  • 作者简介:王珍, E-mail: wangzhen.cq@gmail.com|姚梦楠, E-mail: yaomn3367@163.com
  • 基金资助:
    本研究由国家自然科学基金项目(31872876);本研究由国家自然科学基金项目(31271756);高等学校学科创新引智基地111项目资助(B12006)

Prokaryotic expression, subcellular localization and yeast two-hybrid library screening of BnMAPK1 in B. napus

WANG Zhen(), YAO Meng-Nan(), ZHANG Xiao-Li, QU Cun-Min, LU Kun, LI Jia-Na, LIANG Ying*()   

  1. College of Agronomy and Biotechnology, Southwest University / Chongqing Engineering Research Center for Rapeseed / Academy of Agricultural Sciences, Southwest University, Chongqing 400715, China
  • Received:2020-01-26 Accepted:2020-04-15 Published:2020-09-12 Published online:2020-05-07
  • Contact: Ying LIANG
  • Supported by:
    National Natural Science Foundation of China(31872876);National Natural Science Foundation of China(31271756);Project of Intellectual Base for Discipline Innovation in Colleges and Universities(B12006)

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摘要：

植物的丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinases, MAPKs)级联是生长进程中多种信号跨膜传递的共同通路, 它可以将外源刺激转入细胞内并引发细胞应答。目前C族MAPKs在甘蓝型油菜中的研究报道还很有限。本研究通过1个甘蓝型油菜的C族BnMAPK1基因的生物学进程聚类分析发现, BnMAPK1可能参与蛋白磷酸化、生长素介导的信号途径、逆境应答、细胞周期及转录等过程。BnMAPK1具有369个氨基酸残基, 其相对分子质量约为42.5 kD, 其可溶性蛋白可在原核系统中被诱导表达。BnMAPK1的亚细胞定位结果显示, BnMAPK1主要在细胞核内表达。Bait质粒pGBKT7-BnMAPK1在酵母双杂交系统中无毒性及自激活活性。为深入研究BnMAPK1蛋白参与的生物学进程, 从甘蓝型油菜中油821苗期根、茎、叶中分别提取总RNA, 分离得到mRNA, 利用SMART技术合成并纯化双链cDNA后建立甘蓝型油菜混合cDNA文库。以共转化法筛选与BnMAPK1相互作用的蛋白, 对其分析与鉴定显示, BnMAPK1在生长发育、非生物及生物逆境、转录、蛋白合成及代谢、翻译及翻译后修饰等过程中起作用。研究结果为MAPKs级联, 尤其是C族MAPKs的研究提供了新的视野, 为甘蓝型油菜抗性的机理研究及分子育种奠定了重要的理论依据。

关键词: 甘蓝型油菜, BnMAPK1, 原核表达, 亚细胞定位, 酵母双杂交

Abstract:

Plant mitogen-activated protein kinases (MAPKs) cascades are common signal transduction pathways across the membrane in growth process, they are involved in transducing extracellular stimuli to induce a range of cellular responses. To date, little is known about the role of C-group MAPKs in B. napus. In this study, we analyzed biological process ontology of one C-group BnMAPK1 gene in rapeseed, indicating that BnMAPK1 may be involved in protein phosphorylation, auxin-mediated signaling, stress response, cell cycles and transcription. With 369 amino acid residues, the hydrophilic protein of BnMAPK1 could be induced to express one 42.5 kD protein in the prokaryotic system. The subcellular localization of BnMAPK1 mainly focused in the nucleus. The pGBKT7-BnMAPK1 bait plasmid without toxicity and auto-activation was constructed to perform yeast-two hybrid screening. To further study the function and mechanism of BnMAPK1, we extracted the total RNA of roots, stems, and leaves of B. napus (Zhongyou 821) at seedling stage respectively, and isolated and purified the mRNA to construct the mix cDNA library by SMART method. The BnMAPK1-interacting proteins were screened through co-transformation with bait plasmid and cDNA prey library, demonstrating that BnMAPK1 plays an important role in the growth and development processes, abiotic and biotic stresses, transcription, protein synthesis and metabolism, translation, and post-translational modification, and so on. These results provide a new perspective for the research of MAPKs cascades, especially for C-group MAPKs, and lay a theoretical foundation for the stress resistance mechanism research and molecular breeding in rapeseed.

Key words: Brassica napus, BnMAPK1, prokaryotic expression, subcellular localization, yeast two-hybrid

表1

所用PCR引物"

引物名称
Primer name		 引物序列
Primer sequences (5°-3°)		 用途
Usage
P-BnMAPK1 F: GGATCCATGGCGACACCAGTTGATC
R: GTCGACTAGCAGCTCAGATGTTGA		 BnMAPK1的原核表达载体构建
Construction of prokaryotic expression vector of BnMAPK1
T7 F: TAATACGACTCACTATAGGG
R: TGCTAGTTATTGCTCAGCGG		 原核表达重组载体的检测
Detection of prokaryotic expression recombinant vectors
SC-BnMAPK1 F: ACGAATTCATGGCGACACCAGTTGATC
R: ACGGATCCTAGCAGCTCAGATGTTGA		 BnMAPK1的亚细胞定位表达载体构建
Construction of subcellular localization vector of BnMAPK1
P389 F: ACATGGTCCTGCTGGAGTTC
R: ATTGCCAAATGTTTGAACGA		 亚细胞定位重组载体的检测
Detection of subcellular localization recombinant vectors
eGFP F: ATGGTGAGCAAGGGCGAGGAG
R: GGACTTGTACAGCTCGTCCATGCC		 亚细胞定位重组载体的检测
Detection of subcellular localization recombinant vectors
Y2H-BnMAPK1 F: CGGAATTCATGGCGACACCAGTTGATC
R: CGGGATCCTCATAGCAGCTCAGATGT		 BnMAPK1的酵母双杂交载体构建
Construction of Yeast Two-Hybrid vector of BnMAPK1
BD F: TAATACGACTCACTATAGGG
R: TTTTCGTTTTAAAACCTAAGAGTC		 Bait载体的检测
Detection of Bait vectors
AD F: TAATACGACTCACTATAGGG
R: AGATGGTGCACGATGCACAG		 文库Prey载体的检测
Detection of Prey vectors in library

图1

BnMAPK1的氨基酸组成分析(A)及三维结构预测(N端到C端) (B)"

表2

BnMAPK1的GO生物学进程聚类分析"

序号
No.		 GO编号
GO ID		 GO名称
GO term		 可信度
Reliability (%)
1 GO:0006468 蛋白质氨基酸磷酸化Protein amino acid phosphorylation 46
2 GO:0009734 生长素介导的信号途径Auxin mediated signaling pathway 20
3 GO:0006950 响应逆境Response to stress 18
4 GO:0007049 细胞周期Cell cycle 10
5 GO:0006350 转录Transcription 10

图2

BnMAPK1的蛋白结合位点(A)、二级结构及可溶性(B)和蛋白无序性及构象柔性预测(C)"

图3

BnMAPK1的原核表达(A)、可溶性重组蛋白纯化SDS-PAGE (B)与Branford染色检测(C)"

图4

BnMAPK1-eGFP融合蛋白的亚细胞定位"

图5

总RNA的提取(A)、均一化前(B)后(C)的cDNA及纯化过柱后的cDNA (D)"

图6

Bait-BnMAPK1的毒性和自激活检测"

附图1

BnMAPK1酵母双杂交文库筛选互作蛋白的回转验证"

图7

BnMAPK1候选阳性互作蛋白的调控网络"

附表1

BnMAPK1酵母双杂交文库筛选互作蛋白的基因注释"

编号
No.		 拟南芥基因号
Ath. Gene ID		 油菜基因号
Bna. Gene ID		 出现次数
Frequency		 基因名称
Gene Symbols		 基因注释信息
Description
1 AT3G55980 BnaC08g26910D 12 SZF1 Salt-inducible zinc finger 1
2 AT4G33010 BnaA01g03860D 1 GLDP1 Glycine decarboxylase P-protein 1
3 AT3G10350 BnaA01g31840D 3 GET3B GUIDED ENTRY OF TAIL-ANCHORED PROTEINS 3B//One of 3 GET paralogs in Arabidopsis. GET3b is a chloroplast localized protein with no obvious role in Tail Anchored (TA) protein insertion.
4 AT5G54270 BnaA02g09700D 1 LHCB3 Lhcb3 protein is a component of the main light harvesting chlorophyll a/b-protein complex of Photosystem II (LHC II).
5 AT4G19420 BnaC01g11750D 2 PAE8 Pectin acetylesterase 8//Pectin acetylesterase family protein
6 AT3G05580 BnaA02g30910D 1 TOPP9 Encodes a Type One Protein Phosphatase that acts as a nucleocytoplasmic negative regulator of tip growth. Mutants affect pollen germination, pollen tube growth, and root hair growth. It acts genetically downstream of ANX1 (AT3G04690) and ANX2 (AT5G28680)
7 AT3G62410 BnaA09g39860D 2 CP12-2 CP12-2 encodes a small peptide in the chloroplast stroma. CP12-2 is coordinately regulated by light with the photosynthetic GAPDH and PRK
8 AT4G04770 BnaA09g20260D 2 ABC1, LAF6, ATNAP1 Encodes an iron-stimulated ATPase. A member of the NAP subfamily of ABC transporters. Involved in Fe-S cluster assembly. Similar to SufB. Involved in the regulation of iron homeostasis. Able to form homodimers. Interacts with AtNAP7 inside the chloroplast.
9 AT1G76160 BnaA07g32400D 1 SKS5 SKU5 similar 5
10 AT1G21910 BnaAnng28960D 1 DREB26 encodes a member of the DREB subfamily A-5 of ERF/AP2 transcription factor family. The protein contains one AP2 domain.
11 AT4G25520 BnaA08g30970D 1 SLK1 SEUSS-like 1
12 AT5G28050 BnaA06g29050D 1 GSDA GUANOSINE DEAMINASE, Cytidine/deoxycytidylate deaminase family protein
13 AT2G17040 BnaC03g60970D 1 ANAC036 NAC DOMAIN CONTAINING PROTEIN 36, member of the NAC transcription factor family and more specifically, the ONAC022 subfamily
14 AT2G37130 BnaC03g20530D 1 _ Peroxidase superfamily protein
15 AT3G07020 BnaA05g30520D 1 SGT, UGT80A2 Encodes a 3beta-hydroxy sterol UDP-glucosyltransferase. ugt80a2 mutant plants have reduced steryl glycoside and acyl steryl glycoside levels and reduced seed size. ugt80a2/b1 double mutants have normal levels of celluose and normal cold stress tolerance.
16 AT3G01280 BnaA05g37480D 1 VDAC1 Encodes a voltage-dependent anion channel (VDAC: AT3G01280/VDAC1, AT5G67500/VDAC2, AT5G15090/VDAC3, AT5G57490/VDAC4, AT5G15090/VDAC5). VDACs are reported to be porin-type, beta-barrel diffusion pores. They are prominently localized in the outer mitochondrial membrane and are involved in metabolite exchange between the organelle and the cytosol
17 AT5G67500 BnaA09g07330D 1 VDAC2 Encodes a voltage-dependent anion channel (VDAC: AT3G01280/VDAC1, AT5G67500/VDAC2, AT5G15090/VDAC3, AT5G57490/VDAC4, AT5G15090/VDAC5). VDACs are reported to be porin-type, beta-barrel diffusion pores. They are prominently localized in the outer mitochondrial membrane and are involved in metabolite exchange between the organelle and the cytosol
18 AT5G13180 BnaA10g20110D 1 ANAC083, VNI2 NAC DOMAIN CONTAINING PROTEIN 83, encodes a NAC domain transcription factor that interacts with VND7 and negatively regulates xylem vessel formation
19 AT5G24150 BnaC04g28910D 1 SQP1,SQE5 squalene monooxygenase gene homolog
20 AT5G67030 BnaA07g12170D 1 ABA1, ATZEP, IBS3, LOS6, NPQ2 Encodes a single copy zeaxanthin epoxidase gene that functions in first step of the biosynthesis of the abiotic stress hormone abscisic acid (ABA)
21 AT4G39800 BnaC01g00680D 1 ATIPS1, MIPS1 myo-inositol-1-phosphate synthase
22 AT1G01140 BnaA09g51540D 1 CIPK9, PKS6, SnRK3.12 Encodes a CBL-interacting protein kinase with similarity to SOS2
23 AT2G01110 BnaC02g33640D 1 APG2,UNE3,PGA2,TATC Chloroplast Protein Translocation (tatC), mutant is Albino and pale green. Core subunit of the chloroplast Tat translocase. Integral chloroplast thylakoid membrane protein.
24 AT2G26690 BnaA07g12830D 1 NPF6.2 Major facilitator superfamily protein
25 AT5G61770 BnaA09g05800D 1 PPAN, SNAIL1 A single-copy gene encoding a 346 aa protein with a single Brix domain. Similar to yeast ribosome biogenesis proteins Ssf1/2.
26 AT4G12060 BnaC09g23500D 1 CLPT2 Double Clp-N motif protein
27 AT1G12350 BnaCnng03720D 1 COAB encodes phosphopantothenoylcysteine synthetase (phosphopantothenoylcysteine ligase)
28 AT3G58460 BnaA09g37400D 1 RBL11, RBL15 RHOMBOID-like protein
29 AT5G43940 BnaA09g16810D 1 ADH2, HOT5 Encodes a glutathione-dependent formaldehyde dehydrogenase (also known as class III type alcohol dehydrogenase) reduces S-nitrosoglutathione (GSNO), the condensation product of glutathione and NO, that is a naturally occurring NO reservoir and also a reactive nitrogen intermediate. Gene expression is reduced by wounding and induced by salicylic acid. Is required for the acclimation of plants to high temperature and for fertility
30 AT1G08830 BnaC08g42970D 1 CSD1, SOD1 Encodes a cytosolic copper/zinc superoxide dismutase CSD1 that can detoxify superoxide radicals. Its expression is affected by miR398-directed mRNA cleavage. Regulated by biotic and abiotic stress. Activation of CSD1 in the cytoplasm involves both a CCS-dependent and -independent pathway.
31 AT2G42500 BnaC04g02330D 1 PP2A-3 Encodes one of the isoforms of the catalytic subunit of protein phosphatase 2A: AT1G59830/PP2A-1, AT1G10430/PP2A-2, At2g42500/PP2A-3, At3g58500/PP2A-4 [Plant Molecular Biology (1993) 21:475-485 and (1994) 26:523-528; Note that in more recent publications, there is mixed use of gene names for PP2A-3 and PP2A-4 - some refer to At2g42500 as PP2A-3 and some as PP2A-4]. ACR4 phosphorylates the PROTEIN PHOSPHATASE 2A-3 (PP2A-3) catalytic subunit of the PP2A phosphatase holoenzyme and PP2A dephosphorylates ACR4
32 AT2G02740 BnaC07g21860D 1 PTAC11,WHY3 Encodes a homolog of the potato p24 protein. It shares the conserved KGKAAL domain, a putative DNA-binding domain, with potato p24
33 AT3G19850 BnaC01g32990D 1 _ Phototropic-responsive NPH3 family protein
34 AT2G04880 BnaA03g37950D 1 ZAP1,WRKY1 Encodes WRKY1, a member of the WRKY transcription factors in plants involved in disease resistance, abiotic stress, senescence as well as in some developmental processes. WRKY1 is involved in the salicylic acid signaling pathway. The crystal structure of the WRKY1 C-terminal domain revealed a zinc-binding site and identified the DNA-binding residues of WRKY1.
35 AT3G06080 BnaA03g57390D 1 TBL10 Encodes a member of the TBL (TRICHOME BIREFRINGENCE-LIKE) gene family containing a plant-specific DUF231 (domain of unknown function) domain.
36 AT5G23120 BnaC07g49760D 2 HCF136 encodes a stability and/or assembly factor of photosystem II
37 AT1G53210 BnaCnng29300D 1 NCL Encodes a Na+/Ca 2+ exchanger-like protein that participates in the maintenance of Ca 2+ homeostasis
38 AT5G60210 BnaA02g06460D 1 RIP5 Encodes RIP5 (ROP interactive partner 5), a putative Rho protein effector, interacting specifically with the active form of ROPs (Rho proteins of plants)
39 AT3G02850 BnaCnng06010D 1 SKOR Encodes STELAR K+ OUTWARD RECTIFIER, a member of Shaker family potassium ion (K+) channel. Mediates the delivery of K+ from stelar cells to the xylem in the roots towards the shoot. mRNA accumulation is modulated by abscisic acid. K+ gating activity is modulated by external and internal K+. Involved in response to low potassium.
40 AT5G54110 BnaC02g13800D 1 MAMI Encodes a highly polar protein with more than 60% hydrophilic amino acid residues that is associated with the plasma membrane. It has limited secondary structure similarity to VAP-33 from Aplysia, which may be involved in membrane trafficking.
41 AT5G17020 BnaC09g40920D 1 XPO1A, CRM1, HIT2 Encodes a member of the exportin protein family (XPO1A) which functions as receptors for nuclear export. Binds to a variety of proteins having leucine rich export signals.Along with XPO1B involved with development of the male and female gametophytes.
42 AT4G22690 BnaA08g10430D 1 CYP706A1 Cytochrome P450, family 706, subfamily A, polypeptide 1, member of CYP706A
43 AT2G35500 BnaC04g44410D 1 SKL2 Encodes a protein with some sequence similarity to shikimate kinases, but a truncated form of this protein (lacking a putative N-terminal chloroplast transit peptide) does not have shikimate kinase activity in vitro
44 AT2G38740 BnaA03g18060D 1 _ Haloacid dehalogenase-like hydrolase (HAD) superfamily protein
45 AT3G46060 BnaA06g19870D 1 ARA3, RAB8A, RABE1C small GTP-binding protein (ara-3)
46 AT1G13560 BnaC08g40270D 1 AAPT1 Encodes aminoalcoholphosphotransferase AAPT1
47 ATCG01310 BnaC03g31140D 1 RPL2 encodes a chloroplast ribosomal protein L2, a constituent of the large subunit of the ribosomal complex
48 AT2G16900 BnaC07g05930D 1 PEARLI4 phospholipase-like protein (PEARLI 4) family protein
49 AT4G35110 BnaAnng11620D 1 PEARLI 4 phospholipase-like protein (PEARLI 4) family protein
50 AT3G07720 BnaA05g29930D 1 _ Galactose oxidase/kelch repeat superfamily protein
51 AT1G66720 BnaA03g03600D 1 _ S-adenosyl-L-methionine-dependent methyltransferases superfamily protein
52 AT1G08930 BnaC08g42900D 1 ERD6 Early response to dehydration 6, encodes a putative sucrose transporter whose gene expression is induced by dehydration and cold
53 AT5G19510 BnaA02g04260D 1 _ Translation elongation factor EF1B/ribosomal protein S6 family protein
54 AT4G16360 BnaA08g06000D 1 KINβ2 5'-AMP-activated protein kinase beta-2 subunit protein
55 AT4G11150 BnaC03g72900D 1 TUF, EMB2448, VHA-E1 Encodes a vacuolar H+-ATPase subunit E isoform 1 which is required for Golgi organization and vacuole function in embryogenesis
56 AT5G18370 BnaC09g39520D 1 DSC2 Dominant supressor of camta3 number 2, Leucine-rich repeat domain (NLR) receptor. Dominant negative alleles suppress catma3 autoimmunity
57 AT3G13360 BnaC01g37720D 1 WIP3 WPP domain interacting protein 3, encodes an outer nuclear membrane protein that anchors RanGAP1 to the nuclear envelope. It interacts with SUN proteins and is required for maintaining the elongated nuclear shape of epidermal cells.
58 AT5G35630 BnaC08g07930D 1 GS2, GLN2, GSL1 chloroplastic glutamine synthetase
59 AT1G80750 BnaC06g19470D 1 RPL7A Ribosomal protein L30/L7 family protein
60 AT3G50770 BnaCnng69860D 1 CML41 calmodulin-like 4
61 AT4G36250 BnaC03g61970D 1 ALDH3F1 Encodes a putative aldehyde dehydrogenase 3F1. The gene is not responsive to osmotic stress and is expressed constitutively at a low level in plantlets and root cultures.
62 AT5G38660 BnaA07g14540D 1 APE1 Acclimation of photosynthesis to environment, mutant has Altered acclimation responses
63 AT3G62030 BnaA09g39610D 1 ROC4/CYP20-3 Nuclear-encoded chloroplast stromal cyclophilin CYP20-3 (also known as ROC4). Protein is tyrosine-phosphorylated and its phosphorylation state is modulated in response to ABA in Arabidopsis thaliana seeds
64 AT1G60710 chrC09:10,805,646..10,808,849 1 ATB2 Encodes ATB2
65 AT1G12800 BnaA06g07940D 1 SDP S1 domain-containing RBP, SDP is a chloroplast localized RNA binding protein that is required for plastid rRNA processing. Plants harboring a mutation in SDP have numerous defects including reduced chlorophyll content, poor growth, yellow leaves and abnormal chloroplasts.
66 AT4G37470 BnaA03g53980D 1 KAI2 Karrikin insensitive 2, HTL belonging to the alpha/beta fold hydrolase superfamily. Mutant and over-expression studies indicates its involvement in seedling de-etiolation process.
67 AT1G04250 BnaCnng41350D 1 IAA17, AXR3 Transcription regulator acting as repressor of auxin-inducible gene expression. Auxin-inducible AUX/IAA gene. Short-lived nuclear protein with four conserved domains. Domain III has homology to beta alpha alpha dimerization and DNA binding domains. Involved in auxin signaling and is a positive modulator of natural leaf senescence. Auxin induces the degradation of the protein in a dosage-dependent manner in a process mediated by AtRac1. Auxin induced the relocalization of the protein within the nucleus from a diffused nucleoplasmic pattern to a discrete particulated pattern named nuclear protein bodies or NPB in a process also mediated by Rac1. Colocalizes with SCF, CSN and 26S proteasome components.
68 AT4G04640 BnaA03g25540D 1 ATPC1 One of two genes (with ATPC2) encoding the gamma subunit of Arabidopsis chloroplast ATP synthase.
69 AT1G78510 BnaC06g38940D 1 SPS1 Encodes one of the two paralogous solanesyl diphosphate synthases - SPS1 (At1g78510) and SPS2 (At1g17050) - that assemble the side-chain of plastoquinone-9 in plastids.
70 AT4G01940 BnaA03g26440D 1 NFU1 Encodes a protein containing the NFU domain that may be involved in iron-sulfur cluster assembly. Part of a five member gene family, more closely related to NFU2 and 3 than to NFU4 and 5. Targeted to the chloroplast.
71 AT1G23190 BnaC08g06210D 1 PGM3 Encodes a cytosolic phosphoglucomutase 3, loss of both PGM2 and PGM3 severely impairs male and female gametophyte development
72 AT2G05830 BnaA07g04420D 1 MTI1 Encodes a 5-methylthioribose-1-phosphate isomerase( 5-methylthioribose kinase 1)
73 AT2G46820 BnaA05g01060D 1 CURT1B, PSI_P Encodes the P subunit of Photosystem I. About 25% of the TMP14 pool appeared to be phosphorylated, and this ratio is not affected by light. Contains seven phosphorylation sites on threonine residue and chloroplast targeting signal. Located in the proximity of PSI-L, -H and -O subunits.
74 AT1G80840 BnaA07g35260D 1 WRKY40 Pathogen-induced transcription factor. Binds W-box sequences in vitro. Forms protein complexes with itself and with WRKY40 and WRKY60. Coexpression with WRKY18 or WRKY60 made plants more susceptible to both P. syringae and B. cinerea. WRKY18, WRKY40, and WRKY60 have partially redundant roles in response to the hemibiotrophic bacterial pathogen Pseudomonas syringae and the necrotrophic fungal pathogen Botrytis cinerea, with WRKY18 playing a more important role than the other two.
75 AT2G32860 BnaC04g11860D 1 BGLU33 beta glucosidase 33
76 AT1G06650 BnaC08g44860D 1 _ encodes a protein whose sequence is similar to 2-oxoglutarate-dependent dioxygenase
77 AT1G56500 BnaC09g16020D 1 SOQ1 Suppressor of quenching, Encodes a thylakoid membrane protein with thioredoxin-like and beta-propeller domains located in the lumen and a haloacid-dehalogenase domain exposed to the chloroplast stroma. The protein's role may be to prevent formation of a slowly reversible form of antenna quenching, thereby maintaining the efficiency of light harvesting
78 AT1G13450 BnaC05g09890D 1 GT-1 Encodes GT-1, a plant transcription factor that binds to one of the cis-acting elements, BoxII, which resides within the upstream promoter region of light-responsive genes. GT-1 was assumed to act as a molecular switch modulated through Ca(2+)-dependent phosphorylation/dephosphorylation in response to light signals.
79 AT2G44530 BnaA04g25770D 1 PRS5 Phosphoribosyltransferase family protein
80 AT3G24150 BnaC07g07420D 1 KIX8 Kinase-inducible domain interacting 8, hypothetical protein
81 AT5G19180 BnaA10g15880D 1 ECR1 E1 C-Terminal related 1, encodes a subunit of a RUB-activating enzyme analogous to the E1 ubiquitin-activating enzyme. ECR1 functions as a heterodimer with AXR1 to activate RUB, a ubiquitin-related protein
82 AT1G35620 BnaAnng04610D 1 PDI8, PDIL5-2 Encodes a protein disulfide isomerase-like (PDIL) protein, a member of a multigene family within the thioredoxin (TRX) superfamily. Unlike several other PDI family members, transcript levels for this gene are not up-regulated in response to three different chemical inducers of ER stress (dithiothreitol, beta-mercaptoethanol, and tunicamycin). However, the level of transcripts for this gene is slightly elevated in atbzip60 mutants
83 AT1G03905 BnaC08g00400D 1 ABCI19, ATNAP4 ATP-binding cassette I19, non-intrinic ABC protein 4, P-loop containing nucleoside triphosphate hydrolases superfamily protein
84 AT4G17040 BnaA01g17990D 1 CLPR4 Encodes the ClpR4 subunit of the chloroplast-localized Clp protease complex. hon mutations disturb plastid protein homeostasis, thereby activating plastid signaling and inducing stress acclimatization.
85 AT3G22425 BnaC01g31520D 1 HISN5A, IGPD Encodes imidazoleglycerolphosphate dehydratase
86 AT3G11770 BnaC05g41420D 1 RICE1 Encodes a 23kDa protein with 3'-5' exoribonuclease activity. It is expressed ubiquitously and localized to the cytoplasm.
87 AT5G46760 BnaC07g19420D 1 MYC3 MYC3 is a JAZ-interacting transcription factor that act together with MYC2 and MYC4 to activate JA-responses
88 AT5G51970 BnaCnng32750D 1 SDH Encodes a putative sorbitol dehydrogenase that can be thiolated in vitro
89 AT4G24690 BnaC01g16550D 1 NBR1 Encodes NEXT TO BRCA1 GENE 1, a selective autophagy substrate
90 AT5G44190 BnaC07g17710D 1 GLK2, GPRI2 Encodes GBF's pro-rich region-interacting factor 2, Golden2-like 2, one of a pair of partially redundant nuclear transcription factors that regulate chloroplast development in a cell-autonomous manner.
91 AT5G24530 BnaC02g41200D 1 DMR6 Encodes a putative 2OG-Fe(II) oxygenase that is defense-associated but required for susceptibility to downy mildew
92 AT3G59690 BnaC08g29930D 1 IQD13 IQ-domain 13
93 AT5G06600 BnaC09g48960D 1 UBP12 Encodes a ubiquitin-specific protease 12
94 AT1G27370 BnaC03g57620D 1 SPL10 Squamosa promoter binding protein-like 10, In conjunction with SPL11 and SPL2, SPL10 redundantly controls proper development of lateral organs in association with shoot maturation in the reproductive phase. SPL10 also controls lamina shape during vegetative development.
95 AT4G04020 BnaCnng74600D 1 FIB1A, PGL35 Fibrillin precursor protein. The fibrillin preprotein, but not the mature protein interacts with ABI2. Regulated by abscisic acid response regulators. Involved in abscisic acid-mediated photoprotection.
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