Functional Study of BbRho5 on the Growth Rate of Beauveria bassiana

doi:10.13560/j.cnki.biotech.bull.1985.2021-0210

Abstract

Abstract:

In order to clarify the role of BbRho5 in the biocontrol potential of B. bassiana,a Bbrho5 single gene knockout strain ΔBbrho5 was constructed. The colony growth rate was measured on different media while the wild-type strain WT was used as the control. In addition,carbendazim-tolerance of WT and ΔBbrho5 and their virulence to Galleria mellonella larval body wall were also tested. The transcriptome data of ΔBbrho5 and WT were further obtained and analyzed. The results showed that the functional defect of BbRho5 protein significantly inhibited the hyphal growth rate of B. bassiana;meanwhile BbRho5 showed mild effect in carbendazim-tolerance and virulence in B. bassiana. Compared with WT,there were 770 differentially expressed genes（DEGs）in ΔBbrho5,of which 395 were up-regulated and 375 were down-regulated. GO analysis showed that DEGs in ΔBbrho5 VS WT were mainly enriched in oxidoreductase activity and monooxygenase activity. KEGG pathway enrichment showed that DEGs were mainly enriched in nitrogen metabolism and various amino acid metabolism pathways. Seven functional genes were enriched in the nitrogen metabolism pathway,of which 5 were up-regulated and 2 were down-regulated,indicating that ΔBbrho5 enhanced nitrogen source utilization and glutamate synthesis to cope with the growth retardation caused by Bbrho5 deficiency. The above results reveal that the small GTPases BbRho5 has an important effect on the growth rate of B. bassiana,and nitrogen metabolism and amino acid metabolism may be important metabolic pathways.

Key words: small GTPase, BbRha5, Beauveria bassiana, growth rate, transcriptome

GUAN Yi, LI Xin, WANG Ding-yi, DU Xi, ZHANG Long-bin, YE Xiu-yun. Functional Study of BbRho5 on the Growth Rate of Beauveria bassiana[J]. Biotechnology Bulletin, 2022, 38(2): 132-140.

Figures/Tables 10

References 35

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引物名称 Primer	序列 Sequence（5'-3'）
rho5-up-F	AAAAAGGAATTCCGTCGCCAACATCTCCATC
rho5-up-R	AAAAACGGGATCCGTCGTCGGTGGTGGTGAG
rho5-dn-F	AACGTCGACCCATGGCTCGAGTGGTATCCGCC- CCCTTTCT
rho5-dn-R	CGTTAACACTAGTCAGATCCGGTTAGGGAGGC- TTCGTA
rho5-comp-F	ATCCGTCGACCTGCAGCCACGTCGCCAACATCT- CCAT
rho5-comp -R	ACACTAGTCAGATCTTCTAGCAGTGGTGCTCAG- CAGACAA

引物名称 Primer	序列 Sequence（5'-3'）
rho5-up-F	AAAAAGGAATTCCGTCGCCAACATCTCCATC
rho5-up-R	AAAAACGGGATCCGTCGTCGGTGGTGGTGAG
rho5-dn-F	AACGTCGACCCATGGCTCGAGTGGTATCCGCC- CCCTTTCT
rho5-dn-R	CGTTAACACTAGTCAGATCCGGTTAGGGAGGC- TTCGTA
rho5-comp-F	ATCCGTCGACCTGCAGCCACGTCGCCAACATCT- CCAT
rho5-comp -R	ACACTAGTCAGATCTTCTAGCAGTGGTGCTCAG- CAGACAA

Sample	Clean reads	Clean bases	Error rate/%	Q20/%	Q30/%	GC content/%
WT-1	54944640	8177521548	0.023	98.86	96.11	55.56
WT-2	54680452	8142825538	0.0233	98.76	95.82	55.61
WT-3	53371986	7953327629	0.0234	98.73	95.75	55.71
ΔBbrho5-1	50282602	7496483802	0.0233	98.74	95.81	56.01
ΔBbrho5-2	49584208	7395969389	0.0234	98.75	95.77	55.71
ΔBbrho5-3	52574486	7847183298	0.0232	98.82	96	55.94

Sample	Clean reads	Clean bases	Error rate/%	Q20/%	Q30/%	GC content/%
WT-1	54944640	8177521548	0.023	98.86	96.11	55.56
WT-2	54680452	8142825538	0.0233	98.76	95.82	55.61
WT-3	53371986	7953327629	0.0234	98.73	95.75	55.71
ΔBbrho5-1	50282602	7496483802	0.0233	98.74	95.81	56.01
ΔBbrho5-2	49584208	7395969389	0.0234	98.75	95.77	55.71
ΔBbrho5-3	52574486	7847183298	0.0232	98.82	96	55.94

Sample	Total reads	Total mapped	Multiple mapped	Uniquely mapped
WT-1	54944640	52334433（95.25%）	242381（0.44%）	52092052（94.81%）
WT-2	54680452	52109660（95.3%）	225669（0.41%）	51883991（94.89%）
WT-3	53371986	50353743（94.34%）	210188（0.39%）	50143555（93.95%）
ΔBbrho5-1	50282602	47863412（95.19%）	202423（0.4%）	47660989（94.79%）
ΔBbrho5-2	49584208	47574991（95.95%）	239083（0.48%）	47335908（95.47%）
ΔBbrho5-3	52574486	50041097（95.18%）	302786（0.58%）	49738311（94.61%）