抗、感品种棉花根系分泌物对尖孢镰刀菌生长及基因表达的影响

doi:10.13560/j.cnki.biotech.bull.1985.2023-0284

摘要/Abstract

摘要：

为探究抗、感品种棉花根系分泌物对尖孢镰刀菌生长及基因表达的影响，以棉花感病品种新海14号（XH-14）和抗病品种新海41号（XH-41）为材料，将棉花根系分泌物与尖孢镰刀菌（F327）共培养，观察尖孢镰刀菌的生长；取培养0、6、12、24和48 h样本，进行RNA-seq。结果表明，与XH-41根系分泌物处理的菌落相比，XH-14根系分泌物处理的菌落生长速率快、孢子萌发率高、产孢量显著增加。GO（gene ontology）分析发现，XH-14根系分泌物处理的差异表达基因（differentialy expressed genes, DEGs）主要富集在氨基酸转运、脂肪酸生物合成过程、膜的组成部分、离子通道活性；XH-41根系分泌物处理的DEGs主要富集在跨膜转运、三羧酸循环、膜的组成部分、ATP酶活性。KEGG（Kyoto encyclopedia of genes and genomes）分析发现，XH-14根系分泌物处理的DEGs主要富集在TCA循环、类胡萝卜素生物合成、谷胱甘肽代谢、核黄素代谢等代谢通路；XH-41根系分泌物处理的主要DEGs富集在碳代谢、其他聚糖降解、脂肪酸代谢、ABC运输工具等代谢通路。综合分析结果，获得24个与细胞壁降解酶相关的基因，其中感病品种棉花根系分泌物处理6 h下“水解酶活性、水解O-糖基化合物”基因富集数目最多。研究结果为筛选棉花枯萎病菌潜在的关键致病基因奠定基础，为棉花枯萎病的防控提供理论依据。

关键词: 棉花, 根系分泌物, 尖孢镰刀菌, RNA-seq, 枯萎病, 差异表达基因, 细胞壁降解酶

Abstract:

This work aims to investigate the effects of root secretions in resistant and disease-susceptible varieties of cotton on the growths and gene expressions of Fusarium oxysporum. The cotton susceptible variety Xinhai 14（XH-14）and the disease-resistant variety Xinhai 41（XH-41）were used as materials, and the cotton root secretion and Fusarium oxysporum（F327）was co-cultured to observe the growth of the bacteria; and the samples were taken at 0, 6, 12, 24 and 48 h of incubation and RNA-seq was performed. The results showed that the colonies treated with the XH-14 root exudates grew faster, their spore germination rate was higher, and the sporulation significantly increased, when compared with the colonies treated with XH-41 root secretion. GO（Gene Ontology）analysis revealed that the differentially expressed genes（DEGs）of colonies treated with XH-14 root secretion were mainly enriched in amino acid transport, fatty acid biosynthetic process, membrane components, and ion channel activity. The DEGs under XH-41 root secretion treatment were mainly enriched in transmembrane transport, tricarboxylic acid cycle, membrane components, and ATPase activity. KEGG（Kyoto Encyclopedia of Genes and Genomes）analysis uncovered that DEGs under XH-14 root secretion treatment were were mainly enriched in metabolic pathways such as TCA cycle, carotenoid biosynthesis, glutathione metabolism and riboflavin metabolism. The main DEGs under XH-41 root secretion treatment were enriched in metabolic pathways such as carbon metabolism, other glycan degradation, fatty acid metabolism and ABC transporters. The results of the comprehensive analysis showed that 24 genes related to cell wall degradation enzymes were obtained, among which the largest number of genes were enriched in “hydrolase activity, hydrolysis of O-glycosyl compounds” under 6 h of root secretion treatment in the susceptible cotton varieties. The results of this study provide a basis for screening potential key pathogenic genes of cotton wilt fungus and provide a theoretical basis for the prevention and control of cotton wilt.

Key words: cotton, root exudates, Fusarium oxysporum, RNA-seq, Fusarium wilt, differentially expressed genes, cell wall degrading enzyme

娄慧, 朱金成, 杨洋, 张薇. 抗、感品种棉花根系分泌物对尖孢镰刀菌生长及基因表达的影响[J]. 生物技术通报, 2023, 39(9): 156-167.

LOU Hui, ZHU Jin-cheng, YANG Yang, ZHANG Wei. Effects of Root Exudates in Resistant and Susceptible Varieties of Cotton on the Growths and Gene Expressions of Fusarium oxysporum[J]. Biotechnology Bulletin, 2023, 39(9): 156-167.

图/表 10

参考文献 29

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基因名称 Gene name	引物序列Primer sequence（5'-3'）
FOTG_00376	TGCAGCCCTCATCCTTGC（F）
FOTG_00376	ACAGCGCGGATGGTATCG（R）
FOTG_00798	AGCTGCAATCCCACGCTT（F）
FOTG_00798	AAAAGCAAAGGCAGCGGC（R）
FOTG_00997	CCAGCGAGTTCTCGACCC（F）
FOTG_00997	CAGAGAAGGGCCGCCAAA（R）
FOTG_01540	TCTCCAGGGGCCAGTGTT（F）
FOTG_01540	ACTTTGGCCCGATCGCAA（R）
FOTG_02351	CTTCCCAGGCGTGGCTTT（F）
FOTG_02351	GTCAAACAGGGCGAGCCT（R）
FOTG_05128	GCCTCATCACCACCCGTC（F）
FOTG_05128	GCGAGACCAGGTGACCAC（R）
FOTG_05209	AAGACCGGTGCTTCTGCC（F）
FOTG_05209	TGGAGGCGGAGAAGACGA（R）
FOTG_06306	TCAGCCCTGCCGATGTTG（F）
FOTG_06306	GTCGAGACCGAGCAGCTC（R）
FOTG_07131	TGGCTCCAAGCTCTGTGC（F）
FOTG_07131	CTGAGTTTCCGTGGCCGT（R）
FOTG_08398	TGCTTGGTATGGGCGGTG（F）
FOTG_08398	CAGTGCCCAGCTTTCGGA（R）
FOTG_16221	CCCCAGGAGGCCTCAGAT（F）
FOTG_16221	AGCTCCCGCAAACCTGAC（R）
FOTG_18094	ACCACCATCCGCAACCTG（F）
FOTG_18094	AGGTCCCGTTCGTTGCTG（R）

基因名称 Gene name	引物序列Primer sequence（5'-3'）
FOTG_00376	TGCAGCCCTCATCCTTGC（F）
FOTG_00376	ACAGCGCGGATGGTATCG（R）
FOTG_00798	AGCTGCAATCCCACGCTT（F）
FOTG_00798	AAAAGCAAAGGCAGCGGC（R）
FOTG_00997	CCAGCGAGTTCTCGACCC（F）
FOTG_00997	CAGAGAAGGGCCGCCAAA（R）
FOTG_01540	TCTCCAGGGGCCAGTGTT（F）
FOTG_01540	ACTTTGGCCCGATCGCAA（R）
FOTG_02351	CTTCCCAGGCGTGGCTTT（F）
FOTG_02351	GTCAAACAGGGCGAGCCT（R）
FOTG_05128	GCCTCATCACCACCCGTC（F）
FOTG_05128	GCGAGACCAGGTGACCAC（R）
FOTG_05209	AAGACCGGTGCTTCTGCC（F）
FOTG_05209	TGGAGGCGGAGAAGACGA（R）
FOTG_06306	TCAGCCCTGCCGATGTTG（F）
FOTG_06306	GTCGAGACCGAGCAGCTC（R）
FOTG_07131	TGGCTCCAAGCTCTGTGC（F）
FOTG_07131	CTGAGTTTCCGTGGCCGT（R）
FOTG_08398	TGCTTGGTATGGGCGGTG（F）
FOTG_08398	CAGTGCCCAGCTTTCGGA（R）
FOTG_16221	CCCCAGGAGGCCTCAGAT（F）
FOTG_16221	AGCTCCCGCAAACCTGAC（R）
FOTG_18094	ACCACCATCCGCAACCTG（F）
FOTG_18094	AGGTCCCGTTCGTTGCTG（R）

样品名称Sample ID	质控数据Clean reads	原始数据Raw reads	GC/%	Q20/%	Q30/%	参考基因组比对Mapped reads
F0a	23 634 226	7 078 961 596	52.96	98.65	95.81	44 807 027（94.79%）
F0b	24 801 113	7 426 874 552	52.97	98.43	95.31	46 773 111（94.30%）
G12a	19 061 986	5 703 668 454	52.30	97.12	92.43	33 826 843（88.73%）
G12b	22 878 897	6 847 717 368	52.37	98.49	95.48	41 697 513（91.13%）
G24a	23 252 280	6 960 709 832	52.42	98.34	95.12	42 523 192（91.44%）
G24b	19 547 876	5 853 299 798	52.40	97.09	92.38	35 197 149（90.03%）
G48a	28 294 193	8 474 124 618	52.58	98.22	94.77	51 256 558（90.58%）
G48b	23 044 105	6 899 543 008	52.51	97.29	92.79	41 817 664（90.73%）
G6a	25 066 610	7 505 211 560	52.68	98.33	95.05	45 107 683（89.98%）
G6b	22 483 818	6 732 866 920	52.70	98.51	95.51	40 929 019（91.02%）
K12a	20 781 035	6 222 244 284	52.26	96.85	91.94	37 148 780（89.38%）
K12b	20 396 915	6 107 771 748	52.20	97.50	93.22	36 813 504（90.24%）
K24a	22 395 810	6 701 578 958	52.30	97.46	93.20	39 332 014（87.81%）
K24b	19 324 066	5 781 190 570	52.20	97.49	93.24	33 512 754（86.71%）
K48a	20 253 195	6 063 580 670	52.59	97.32	92.80	37 310 755（92.11%）
K48b	23 431 421	7 015 769 462	52.54	98.63	95.77	42 942 424（91.63%）
K6a	19 328 402	5 788 003 554	52.75	97.24	92.64	34 813 490（90.06%）
K6b	20 460 546	6 123 145 102	52.66	97.52	93.25	36 420 714（89.00%）
W12a	20 317 585	6 085 191 946	52.29	98.25	94.84	37 516 540（92.33%）
W12b	23 399 026	7 007 306 032	52.23	98.25	94.86	42 862 234（91.59%）
W24a	22 727 561	6 805 789 076	52.39	98.09	94.46	42 220 799（92.88%）
W24b	21 749 990	6 509 670 558	52.28	98.27	94.83	40 299 468（92.64%）
W48a	26 212 102	7 848 652 478	52.73	98.42	95.30	49 390 772（94.21%）
W48b	25 173 938	7 534 704 604	52.75	97.93	94.12	47 156 136（93.66%）
W6a	22 480 503	6 733 786 866	52.67	98.44	95.33	41 135 869（91.49%）
W6b	26 838 538	8 034 600 906	52.81	98.48	95.43	49 740 879（92.67%）

样品名称Sample ID	质控数据Clean reads	原始数据Raw reads	GC/%	Q20/%	Q30/%	参考基因组比对Mapped reads
F0a	23 634 226	7 078 961 596	52.96	98.65	95.81	44 807 027（94.79%）
F0b	24 801 113	7 426 874 552	52.97	98.43	95.31	46 773 111（94.30%）
G12a	19 061 986	5 703 668 454	52.30	97.12	92.43	33 826 843（88.73%）
G12b	22 878 897	6 847 717 368	52.37	98.49	95.48	41 697 513（91.13%）
G24a	23 252 280	6 960 709 832	52.42	98.34	95.12	42 523 192（91.44%）
G24b	19 547 876	5 853 299 798	52.40	97.09	92.38	35 197 149（90.03%）
G48a	28 294 193	8 474 124 618	52.58	98.22	94.77	51 256 558（90.58%）
G48b	23 044 105	6 899 543 008	52.51	97.29	92.79	41 817 664（90.73%）
G6a	25 066 610	7 505 211 560	52.68	98.33	95.05	45 107 683（89.98%）
G6b	22 483 818	6 732 866 920	52.70	98.51	95.51	40 929 019（91.02%）
K12a	20 781 035	6 222 244 284	52.26	96.85	91.94	37 148 780（89.38%）
K12b	20 396 915	6 107 771 748	52.20	97.50	93.22	36 813 504（90.24%）
K24a	22 395 810	6 701 578 958	52.30	97.46	93.20	39 332 014（87.81%）
K24b	19 324 066	5 781 190 570	52.20	97.49	93.24	33 512 754（86.71%）
K48a	20 253 195	6 063 580 670	52.59	97.32	92.80	37 310 755（92.11%）
K48b	23 431 421	7 015 769 462	52.54	98.63	95.77	42 942 424（91.63%）
K6a	19 328 402	5 788 003 554	52.75	97.24	92.64	34 813 490（90.06%）
K6b	20 460 546	6 123 145 102	52.66	97.52	93.25	36 420 714（89.00%）
W12a	20 317 585	6 085 191 946	52.29	98.25	94.84	37 516 540（92.33%）
W12b	23 399 026	7 007 306 032	52.23	98.25	94.86	42 862 234（91.59%）
W24a	22 727 561	6 805 789 076	52.39	98.09	94.46	42 220 799（92.88%）
W24b	21 749 990	6 509 670 558	52.28	98.27	94.83	40 299 468（92.64%）
W48a	26 212 102	7 848 652 478	52.73	98.42	95.30	49 390 772（94.21%）
W48b	25 173 938	7 534 704 604	52.75	97.93	94.12	47 156 136（93.66%）
W6a	22 480 503	6 733 786 866	52.67	98.44	95.33	41 135 869（91.49%）
W6b	26 838 538	8 034 600 906	52.81	98.48	95.43	49 740 879（92.67%）