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

doi:10.3724/SP.J.1006.2020.04019

Abstract

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

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.

Figures/Tables 11

Table 1

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Supplementary table 1

Genes information of BnMAPK1-interacting proteins in Y2H screening assays"

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.

Supplementary table 1

References 47

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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

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

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

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