Molecular Mechanism of MpICE1 Overexpressing Banana Resistant to Fusarium oxysporum Based on Transcriptome Analysis

doi:10.16420/j.issn.0513-353x.2022-0682

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (10): 2242-2256.doi: 10.16420/j.issn.0513-353x.2022-0682

• Plant Protection • Previous Articles Next Articles

Molecular Mechanism of MpICE1 Overexpressing Banana Resistant to Fusarium oxysporum Based on Transcriptome Analysis

SHI Jingfang¹^,³, HU Chunhua¹, LI Haochen¹, YANG Qiaosong¹^,⁴, SHENG Ou¹, BI Fangcheng¹^,², DONG Tao¹, LI Chunyu¹, DENG Guiming¹, GAO Huijun¹, HE Weidi¹, LIU Siwen¹, YI Ganjun¹^,^*(), DOU Tongxin¹^,^*()

¹ Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization，Ministry of Agriculture and Rural Affairs，Guangdong Key Laboratory of Tropical and Subtropical Fruit Tree Research，Institute of Fruit Tree Research，Guangdong Academy of Agricultural Sciences，Guangzhou 510640，China
² Labortatory of Lingnan Modern Agriculture Project，Guangdong 510642，China
³ Guangdong Provincial Key Laboratory for Crop Germplasm Resources Preservation and Utilization，Agro-Biological Gene Research Center，Guangdong Academy of Agricultural Sciences，Guangzhou 510640，China
⁴ Maoming Branch，Guangdong Labortatory for Lingnan Modern Agriculture，Maoming，Guangdong 525000，China

Received:2023-03-21 Revised:2023-08-13 Online:2023-10-25 Published:2023-10-31

Abstract

Abstract:

Based on previous studies，it was found that overexpression of MpICE1 transcription factor in bananas significantly improved the resistance to fusarium wilt. To further reveal the resistance mechanism of MpICE1 to fusarium wilt，the samples from the roots of wild-type and MpICE1 overexpressing bananas before（0 d）and after （7 d and 14 d）Foc TR4 inoculation were collected， and these samples were applied to RNA-sequencing for DEGs analysis. The results showed that the overexpression of MpICE1 could increase the expression of a series of genes before inoculation. These genes were enriched in phenylpropane synthesis，flavonoid synthesis，ABC transporters，MAPK signal transduction，pentose and glucuronic acid conversion pathway，which were related to the structure and function of cell wall. Combined with KEGG pathway analysis，we speculate that overexpression of MpICE1 may elevate the resistance to Foc TR4 by enhancing the basic immunity. In addition，genes higherly expressed in MpICE1 overexpressing bananas began to decline after inoculation，and the interaction between MpICE1 overexpressing bananas and pathogens reached a relatively stable state seven days after inoculation. Compared with the wild type，only a small amount of DEGs were found in MpICE1 overexpressing bananas 7 and 14 days after incolation. The expression dynamic of DEGs revealed that 12-oxophytodienoic acid reductase （Ma06_g18840），which is related to oxidation lipid metabolism （JA and OPDA），may be the target gene of MpICE1. Moreover，the expression of ubiquitin carboxyl-terminal hydrolase 36/42 was also affected by MpICE1 overexpression. However，the specific mechanism was still unknown，and need further study.

Key words: banana, Fusarium oxysporum, MpICE1, RNA-seq

SHI Jingfang, HU Chunhua, LI Haochen, YANG Qiaosong, SHENG Ou, BI Fangcheng, DONG Tao, LI Chunyu, DENG Guiming, GAO Huijun, HE Weidi, LIU Siwen, YI Ganjun, DOU Tongxin. Molecular Mechanism of MpICE1 Overexpressing Banana Resistant to Fusarium oxysporum Based on Transcriptome Analysis[J]. Acta Horticulturae Sinica, 2023, 50(10): 2242-2256.

Figures/Tables 10

References 43

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基因 Gene	引物（5′-3′） Primer
Ma06_g18840	F：TATGGCAATGTCCCCCAACC；R：GACCAGTGTCTGAGACGCAA
Ma06_g16670	F：ATCAACGTCGAGATCGGCAA；R：GGCTGAGGCTGATGGGTA
Ma11_g19450	F：ATGCTCTCGGGGATCAATGG；R：TCATGACACTGTATTATCG
Ma04_g40060	F：ATGGCGTCCAAGACCACAAG；R：GCTCTTTTCCTCTTGTTCAAC
Ma10_g15940	F：GTCCTCACCAGCGCCTGCGG；R：CCCCGAGCCCTGCCCCCTCCG
Ma10_g12250	F：TGGGTTTGCCATGGATGTTT；R：AGAACACCAGGGCCTTCCTT
Ma08_g05750	F：AGCGGAGAATGTCATAGGTG；R：TGGATGCTCTGTCCATCTTG
Ma09_g10470	F：ACAGTCGTCAGGAGTCGTGCTT；R：CGCTATCGATTCATGACACTGTA

基因 Gene	引物（5′-3′） Primer
Ma06_g18840	F：TATGGCAATGTCCCCCAACC；R：GACCAGTGTCTGAGACGCAA
Ma06_g16670	F：ATCAACGTCGAGATCGGCAA；R：GGCTGAGGCTGATGGGTA
Ma11_g19450	F：ATGCTCTCGGGGATCAATGG；R：TCATGACACTGTATTATCG
Ma04_g40060	F：ATGGCGTCCAAGACCACAAG；R：GCTCTTTTCCTCTTGTTCAAC
Ma10_g15940	F：GTCCTCACCAGCGCCTGCGG；R：CCCCGAGCCCTGCCCCCTCCG
Ma10_g12250	F：TGGGTTTGCCATGGATGTTT；R：AGAACACCAGGGCCTTCCTT
Ma08_g05750	F：AGCGGAGAATGTCATAGGTG；R：TGGATGCTCTGTCCATCTTG
Ma09_g10470	F：ACAGTCGTCAGGAGTCGTGCTT；R：CGCTATCGATTCATGACACTGTA

样本 Sample	原始序列/Mb Raw reads	过滤后序列/Mb Clean reads	Q20/%	Q30/%	序列比值 Clean reads ratio/%	总基因比对率 Total mapping/%
WT0d A	43.82	42.72	97.26	93.03	97.49	87.00
WT0d B	43.82	42.84	97.22	92.84	97.76	86.83
WT0d C	42.61	41.87	96.76	91.99	98.26	86.20
WT7d A	43.82	43.29	97.12	92.54	98.80	88.15
WT7d B	43.82	43.38	97.03	92.41	98.99	87.82
WT7d C	43.82	42.56	97.15	92.71	97.12	87.81
WT14d A	43.82	42.61	97.19	92.84	97.23	87.19
WT14d B	43.82	43.05	97.22	92.91	98.25	87.56
WT14d C	43.82	42.87	97.38	93.25	97.84	87.58
OE0d A	43.82	42.64	97.17	92.88	97.32	87.11
OE0d B	43.82	43.22	97.12	92.67	98.62	87.54
OE0d C	43.82	43.26	96.99	92.31	98.72	86.53
OE7d A	43.82	43.20	97.19	92.76	98.59	87.44
OE7d B	43.82	42.95	97.31	93.15	98.02	87.17
OE7d C	43.82	42.83	97.02	92.45	97.74	87.59
OE14d A	43.82	42.94	96.97	92.27	97.99	87.17
OE14d B	43.82	42.94	97.00	92.41	97.98	87.41
OE14d C	43.82	43.09	96.99	92.32	98.33	86.77

样本 Sample	原始序列/Mb Raw reads	过滤后序列/Mb Clean reads	Q20/%	Q30/%	序列比值 Clean reads ratio/%	总基因比对率 Total mapping/%
WT0d A	43.82	42.72	97.26	93.03	97.49	87.00
WT0d B	43.82	42.84	97.22	92.84	97.76	86.83
WT0d C	42.61	41.87	96.76	91.99	98.26	86.20
WT7d A	43.82	43.29	97.12	92.54	98.80	88.15
WT7d B	43.82	43.38	97.03	92.41	98.99	87.82
WT7d C	43.82	42.56	97.15	92.71	97.12	87.81
WT14d A	43.82	42.61	97.19	92.84	97.23	87.19
WT14d B	43.82	43.05	97.22	92.91	98.25	87.56
WT14d C	43.82	42.87	97.38	93.25	97.84	87.58
OE0d A	43.82	42.64	97.17	92.88	97.32	87.11
OE0d B	43.82	43.22	97.12	92.67	98.62	87.54
OE0d C	43.82	43.26	96.99	92.31	98.72	86.53
OE7d A	43.82	43.20	97.19	92.76	98.59	87.44
OE7d B	43.82	42.95	97.31	93.15	98.02	87.17
OE7d C	43.82	42.83	97.02	92.45	97.74	87.59
OE14d A	43.82	42.94	96.97	92.27	97.99	87.17
OE14d B	43.82	42.94	97.00	92.41	97.98	87.41
OE14d C	43.82	43.09	96.99	92.32	98.33	86.77

比较组 Pair comparison	GO功能 GO term	基因数量 Gene number	富集率 Rich ratio	Q值 Q value
WT 0 d vs. OE 0 d	细胞外区域Extracellular region	56	0.19	6.84E-19
	细胞壁组织或生物发生Cell wall organization or biogenesis	39	0.16	6.10E-10
	细胞壁组织Cell wall organization	34	0.16	2.23E-08
	表面封装结构组织External encapsulating structure organization	34	0.15	3.20E-08
	细胞壁Cell wall	27	0.18	3.66E-08
	表面封装结构External encapsulating structure	27	0.18	3.66E-08
	糖基水解酶活性Hydrolase activity，acting on glycosyl bonds	41	0.15	3.98E-08
	O-糖基水解酶活性 Hydrolase activity，hydrolyzing O-glycosyl compounds	39	0.15	4.60E-08
	细胞碳水化合物代谢过程Cellular carbohydrate metabolic process	28	0.17	1.20E-07
	碳水化合物代谢过程Carbohydrate metabolic process	59	0.10	1.44E-07
WT 7 d vs. OE 7 d	细胞外区域Extracellular region	18	0.06	1.68E-09
	植物细胞壁组织Plant-type cell wall organization	6	0.13	1.47E-04
	氧化还原酶活性Oxidoreductase activity	25	0.02	2.39E-04
	植物细胞壁组织或生物发生 Plant-type cell wall organization or biogenesis	6	0.10	2.56E-04
	细胞外周Cell periphery	13	0.03	9.29E-03
	细胞壁Cell wall	7	0.05	9.73E-03
	表面封装结构External encapsulating structure	7	0.05	9.73E-03
	细胞壁组织或生物发生Cell wall organization or biogenesis	8	0.03	9.73E-03
	质外体Apoplast	5	0.06	2.32E-02
	细胞壁组织Cell wall organization	7	0.03	2.32E-02
WT 14 d vs. OE 14 d	逆转录转座子核衣壳Retrotransposon nucleocapsid	20	0.40	7.45E-11
	DNA整合DNA integration	23	0.29	5.50E-10
	RNA指导的DNA聚合酶活性RNA-directed DNA polymerase activity	10	0.43	2.49E-05
	碳水化合物代谢过程Carbohydrate metabolic process	57	0.10	3.29E-05
	氧化还原酶活性Oxidoreductase activity	93	0.08	2.83E-04
	细胞外区域Extracellular region	34	0.11	5.48E-04
	DNA聚合酶活性DNA polymerase activity	10	0.30	5.48E-04
	甲基丙二酸半醛脱氢酶活性 Methylmalonate-semialdehyde dehydrogenase（acylating）activity	5	0.63	2.20E-03
	质膜组成成分Integral component of membrane	243	0.06	2.20E-03
	质膜固有成分Intrinsic component of membrane	244	0.06	2.20E-03
	催化活性Catalytic activity	353	0.06	4.09E-03

Molecular Mechanism of MpICE1 Overexpressing Banana Resistant to Fusarium oxysporum Based on Transcriptome Analysis

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比较组 Pair comparison	KEGG 通路 KEGG pathway	基因数量 Gene number	富集率 Rich ratio	Q值 Q value
WT 0 d vs. OE 0 d	苯丙烷合成途径Phenylpropanoid biosynthesis	80	0.11	1.43E-09
	黄酮生物合成Flavonoid biosynthesis	23	0.16	7.14E-05
	氰基氨基酸代谢Cyanoamino acid metabolism	21	0.15	4.09E-04
	ABC转运蛋白ABC transporters	26	0.13	4.45E-04
	油菜素内酯生物合成Brassinosteroid biosynthesis	9	0.27	6.06E-04
	单萜生物合成Monoterpenoid biosynthesis	7	0.32	1.36E-03
	植物MAPK信号途径MAPK signaling pathway-plant	76	0.08	1.85E-03
	戊糖和葡萄糖醛酸酯互换 Pentose and glucuronate interconversions	34	0.10	2.16E-03
	角质、软木脂和蜡质生物合成 Cutin，suberine and wax biosynthesis	13	0.15	4.69E-03
	RNA聚合酶RNA polymerase	40	0.09	8.29E-03
	其他聚糖降解Other glycan degradation	17	0.11	2.11E-02
	异喹啉生物碱生物合成Isoquinoline alkaloid biosynthesis	8	0.16	2.77E-02
WT 7 d vs. OE 7 d	黄酮生物合成Flavonoid biosynthesis	12	0.08	1.23E-06
	类胡萝卜素生物合成Carotenoid biosynthesis	6	0.06	6.80E-03
	苯丙烷合成途径Phenylpropanoid biosynthesis	17	0.02	6.80E-03
	植物—病原菌互作Plant-pathogen interaction	20	0.02	6.80E-03
	ABC转运蛋白ABC transporters	8	0.04	9.06E-03
	植物MAPK信号途径MAPK signaling pathway-plant	20	0.02	9.06E-03
	二萜生物合成Diterpenoid biosynthesis	5	0.05	1.45E-02
WT 14 d vs. OE 14 d	黄酮生物合成Flavonoid biosynthesis	23	0.16	2.03E-04
	淀粉和蔗糖代谢Starch and sucrose metabolism	56	0.10	2.09E-04
	植物MAPK信号途径MAPK signaling pathway-plant	81	0.08	1.52E-03
	ABC转运蛋白ABC transporters	25	0.12	2.80E-03
	苯丙烷合成途径Phenylpropanoid biosynthesis	61	0.09	3.20E-03
	植物—病原菌互作Plant-pathogen interaction	75	0.08	3.20E-03
	Alpha-亚油酸代谢alpha-Linolenic acid metabolism	15	0.14	1.33E-02
	二萜生物合成Diterpenoid biosynthesis	13	0.14	1.85E-02
	芪类、二苯基庚烷和姜酚生物合成 Stilbenoid，diarylheptanoid and gingerol biosynthesis	11	0.15	2.58E-02
	RNA聚合酶RNA polymerase	40	0.09	2.58E-02

比较组 Pair comparison	KEGG通路 KEGG pathway	基因数量 Gene number	富集率 Rich ratio	Q值 Q value
OE 0 d vs. OE 7 d（2 003）	苯丙烷生物合成 Phenylpropanoid biosynthesis	89	0.13	5.35E-10
	二萜生物合成 Diterpenoid biosynthesis	21	0.23	2.83E-06
	植物—病原菌互作 Plant-pathogen interaction	94	0.10	2.83E-06
	植物MAPK信号途径 MAPK signaling pathway - plant	96	0.10	7.54E-06
	RNA聚合酶 RNA polymerase	55	0.12	1.14E-05
	黄酮生物合成 Flavonoid biosynthesis	22	0.15	9.43E-04
	ABC转运蛋白 ABC transporters	26	0.13	2.85E-03
	芪类、二苯基庚烷和姜酚生物合成 Stilbenoid，diarylheptanoid and gingerol biosynthesis	13	0.17	5.75E-03
	角质、软木脂和蜡质生物合成 Cutin，suberine and wax biosynthesis	14	0.16	6.14E-03
	甘露糖类O-聚糖生物合成 Mannose type O-glycan biosynthesis	8	0.22	1.13E-02
WT 0 d vs. WT 7 d（1 145）	光合作用天线蛋白 Photosynthesis-antenna proteins	10	0.29	2.41E-05
	植物激素信号传导 Plant hormone signal transduction	65	0.05	1.08E-02
	淀粉和蔗糖代谢 Starch and sucrose metabolism	34	0.06	2.49E-02
	倍半萜和三萜生物合成 Sesquiterpenoid and triterpenoid biosynthesis	11	0.10	3.60E-02
	甘油酯代谢 Glycerolipid metabolism	17	0.07	4.77E-02
OE 0 d vs. OE7 d & WT 0 d vs. WT 7 d（1 897）	植物昼夜节律 Circadian rhythm-plant	67	0.16	2.43E-13
	泛醌和其他萜—醌生物合成 Ubiquinone and other terpenoid-quinone biosynthesis	14	0.16	7.13E-03
	类胡萝卜素生物合成 Carotenoid biosynthesis	16	0.15	7.13E-03
	花生四烯酸代谢 Arachidonic acid metabolism	13	0.17	8.61E-03
	类固醇生物合成 Steroid biosynthesis	11	0.15	3.71E-02
	淀粉和蔗糖代谢 Starch and sucrose metabolism	48	0.09	3.71E-02
OE 7 d vs. OE 14 d（154）	戊糖和葡萄糖醛酸酯相互转化 Pentose and glucuronate interconversions	6	0.02	9.66E-02
	苯丙烷生物合成 Phenylpropanoid biosynthesis	9	0.01	9.66E-02
	硒化合物代谢 Selenocompound metabolism	3	0.04	1.01E-01
	黄酮和黄酮醇生物合成 Flavone and flavonol biosynthesis	2	0.03	2.78E-01
OE 7 d vs. OE 14 d & WT 7 d vs. WT 14 d（264）	黄酮生物合成 Flavonoid biosynthesis	11	0.07	1.86E-05
	内质网蛋白质加工 Protein processing in endoplasmic reticulum	15	0.02	4.52E-02
	植物—病原菌互作 Plant-pathogen interaction	19	0.02	4.52E-02
WT 7 d vs. WT 14 d（3 407）	植物—病原菌互作 Plant-pathogen interaction	169	0.19	1.45E-11
	植物MAPK信号途径 MAPK signaling pathway-plant	166	0.17	5.42E-09
	苯丙氨酸代谢 Phenylalanine metabolism	37	0.27	1.47E-06
	黄酮生物合成 Flavonoid biosynthesis	34	0.23	2.25E-04
	苯丙烷生物合成 Phenylpropanoid biosynthesis	110	0.16	3.48E-04
	光合作用 Photosynthesis	32	0.23	4.57E-04
	光合作用天线蛋白 Photosynthesis-antenna proteins	12	0.34	2.50E-03
	异喹啉生物碱生物合成 Isoquinoline alkaloid biosynthesis	14	0.29	5.82E-03
	谷胱甘肽代谢 Glutathione metabolism	30	0.20	6.99E-03
	芪类、二苯基庚烷和姜酚生物合成 Stilbenoid，diarylheptanoid and gingerol biosynthesis	18	0.24	8.25E-03
	ABC转运蛋白 ABC transporters	37	0.18	9.11E-03
	光合生物碳固定 Carbon fixation in photosynthetic organisms	33	0.18	1.43E-02
	淀粉和蔗糖代谢 Starch and sucrose metabolism	80	0.14	2.84E-02
	黄酮和黄酮醇生物合成 Flavone and flavonol biosynthesis	14	0.23	4.18E-02
	精氨酸生物合成 Arginine biosynthesis	18	0.20	4.37E-02
	酪氨酸代谢 Tyrosine metabolism	16	0.21	4.92E-02

基因 Gene ID	基因表达量（FPKM）						基因功能注释 Gene function annotation
	野生型 Wild type			转基因香蕉 DX11 Transgenic bananas
	0 d	7 d	14 d	0 d	7 d	14 d
Ma11_g19450	8.72	7.66	125.14	645.73	85.73	76.26	泛素羧基末端水解酶36/42 Ubiquitin carboxyl- terminal hydrolase 36/42
Ma10_g12250	0.48	0.76	0.29	491.73	326.85	317.63	转录因子ICE1 Transcription factor ICE1
Ma06_g16670	52.83	15.99	30.77	490.96	79.46	12.19	甘露糖特异类凝集素Mannose-specific lectin-like
Ma04_g33410	2.01	0.98	3.80	220.36	19.60	0.60	泛素羧基末端水解酶36/42 Ubiquitin carboxyl- terminal hydrolase 36/42
Ma11_g22720	11.51	35.87	52.02	179.83	117.17	27.65	类金属硫蛋白2型Metallothionein-like protein type 2
Ma06_g03820	1.65	7.76	82.79	170.87	45.08	16.56	类EXORDIUM 蛋白2 Protein EXORDIUM-like 2
Ma10_g15940	48.26	55.42	226.86	145.97	240.12	88.15	过氧化物酶 Peroxidase
Ma04_g40060	17.93	38.03	16.79	128.60	196.36	51.97	果胶酯酶 Pectinesterase
Ma02_g16400	9.11	11.60	36.76	73.16	60.69	34.71	非特异性脂转运蛋白2 Non-specific lipid-transfer protein 2-like
Ma00_g02560	4.43	0.72	2.53	53.73	3.23	0.62	UDP-葡萄糖醛酸酯4-异构酶 UDP-glucuronate 4-epimerase
Ma04_g31200	0.05	0.10	15.01	52.53	2.20	0.05	果胶酯酶Pectinesterase
Ma03_g33000	8.73	12.50	108.45	39.39	43.65	36.67	DNA指导的RNA聚合酶Ⅱ亚基RPB1 DNA- directed RNA polymerase II subunit RPB1
Ma10 g01930	5.74	4.03	9.69	38.51	14.91	4.65	1-脱氧-D-木酮糖-5-磷酸合成酶 1-Deoxy-D-xylulose- 5-phosphate synthase
Ma09_g10470	3.11	1.51	16.58	37.15	23.73	27.74	类GOS9蛋白Protein GOS9-like
Ma08_g31490	2.09	0.71	1.19	36.48	4.28	1.23	富半胱氨酸和甘氨酸蛋白 Cysteine and glycine-rich protein
Ma09_g25000	9.62	9.09	9.12	35.19	44.91	19.02	糖转运蛋白C Sugar carrier protein C
Ma08_g05750	5.34	18.13	205.01	33.95	91.41	47.89	赤霉素受体GID1 Gibberellin receptor GID1
Ma08_g09150	1.01	0.91	8.82	32.42	11.90	6.70	多酚氧化酶polyphenol oxidase
Ma02 g13020	7.09	4.53	617.71	30.60	30.74	49.95	磷脂：二酰甘油酰基转移酶 Phospholipid：diacylglycerol acyltransferase
Ma03_g28020	8.34	2.91	159.63	25.75	10.80	11.54	几丁质酶1亚型X1 Chitinase 1 isoform X1
Ma11_g25080	5.53	22.31	77.52	19.21	114.19	48.29	糖转运家族32 Solute carrier family 32
Ma08_g12790	0.09	1.53	63.49	19.07	97.90	34.95	DNA连接酶1 DNA ligase 1
Ma09_g20710	1.58	2.09	194.79	11.40	20.77	12.12	几丁质酶 chitinase
Ma09 g11910	3.01	2.83	30.79	12.41	9.83	5.99	白介素-1受体相关激酶4 Interleukin-1 receptor- associated kinase 4
Ma04_g09070	0.00	0.04	85.83	13.85	5.26	0.12	糖转运家族50 Solute carrier family 50
Ma06_g31980	1.13	0.99	124.83	3.44	19.23	6.20	几丁质酶Chitinase
Ma11_g21980	0.08	0.16	35.66	3.66	15.75	0.08	扩张蛋白Expansin
Ma00_g04060	0.12	0.12	0.11	3.61	2.23	0.57	丝裂原激活蛋白激酶激酶4/5 Mitogen-activated protein kinase kinase 4/5
Ma01_g04570	0.00	0	0.05	3.49	1.95	0.16	丝裂原激活蛋白激酶激酶4/5 Mitogen-activated protein kinase kinase 4/5
Ma06_g18840	21.60	109.27	228.42	0.08	0.28	0.20	12-氧-植物二烯酸氧化还原酶 12-Oxophytodienoic acid reductase

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