Study on the Expression Pattern of Genes in Lignin Biosynthesis Pathway of Cassava Resisting to Tetranychus urticae

doi:10.13560/j.cnki.biotech.bull.1985.2022-0500

Abstract

Abstract:

Lignin is an important physicochemical barrier for crops to defend against pests, however, its mechanism of cassava resistance to insect remains unclear. In order to explore the role of lignin biosynthesis pathway genes in cassava resisting to Tetranychus urticae（two-spotted spider mite, TSSM）, the mite-resistant cassava cultivar C1115, Myanmar, SC9 and mite-susceptible cassava cultivar SC205, Bread and BRA900 were applied during mite-cassava interaction. Furthermore, real-time quantitative PCR was used to analyze the expression pattern of lignin biosynthesis pathway genes（PAL, 4CL, C4H, HCT, CSE, COMT, CCoAOMT, F5H, CCR and CAD）among different cultivars infested by TSSM at different times of 0, 1, and 4 d. The results showed that the expressions of C4H, HCT, CSE, F5H and CAD in TSSM-infested mite-resistant and mite-susceptible cassava cultivars generally increased compared with those before infestation, but they did not always show significant differences, in addition, there was no significant difference in the expressions of these five genes among the mite-resistant and mite-susceptible cassava cultivars at the identical TSSM infested time. On the contrary, compared with those before infestation, the expressions of PAL, 4CL, CCoAOMT and CCR significantly increased in the TSSM infested mite-resistant cassava cultivars. However, the expressions of those four genes in mite-susceptible cultivars were either unchanged or significantly decreased, moreover, during the same mite-infested time, the expressions of these four genes in mite-resistant cassava cultivars were generally higher than those in mite-infested cassava cultivars, especially when the cassava plants were infested by TSSM at 4 d. Further correlation analysis found that the increase in the expressions of these four genes was significantly and positively correlated with the resistance of cassava to mite. To sum up, the above results speculated that the expressions of lignin biosynthesis pathway genes PAL, 4CL, CCoAOMT and CCR may be probably related to the resistance level of cassava cultivars.

Key words: cassava（Manihot esculenta Crantz）, Tetranychus urticae, lignin biosynthesis pathway, gene expression, mite-resistant and mite-susceptible cassava cultivars, mite infestation time, correlation of resistance

YAO Xiao-wen, LIANG Xiao, CHEN Qing, WU Chun-ling, LIU Ying, LIU Xiao-qiang, SHUI Jun, QIAO Yang, MAO Yi-ming, CHEN Yin-hua, ZHANG Yin-dong. Study on the Expression Pattern of Genes in Lignin Biosynthesis Pathway of Cassava Resisting to Tetranychus urticae[J]. Biotechnology Bulletin, 2023, 39(2): 161-171.

Figures/Tables 11

Table 1 Candidate gene primers in the synthesis pathways of cassava（Manihot esculenta Crantz）lignin

Gene name	Gene ID	Primer sequence（5'-3'）	Tm/℃	Product size/bp
PAL	MANES_08G008400	F: AGGCATTTGGAGGAGAACTTGA R: TCGCAGAATCTTGATGGGTGA	61.2 59.8	113
C4H	MANES_18G126900	F: GCTCGCCAACAATCCTGCTC R: CTCCTCCTTCCAACTCCAAACG	58.7 59.3	158
4CL	MANES_17G006400	F: GGACTGTTGTCACTGGGCACTA R: TCTTCGTCAACGCTTGGAGTA	59.9 58.5	133
CCR	MANES_04G103200	F: GTCAATGCTGTTCAAGGCTATGTG R: AAATGTATCGCCCGGAGGC	61.6 62.2	94
CAD	MANES_13G117800	F: ACCAGAGGGAATGTCACCAGAA R: CCCTACTCCTCCAAGCCCTAAT	60.8 60.9	133
HCT	MANES_04G101700	F: GACCTTGTGGTGCCGAGATTC R: CGTCCAGCCATCGGATAGAAC	61.7 61.9	137
CSE	MANES_12G157800	F: TCCAATCGGAACCCAACACG R: AAGAGCAGCCCATACACGAAGAG	63.6 62.7	112
CCoAOMT	MANES_10G078800	F: GCCCACATCAATGGCTTCC R: GCTCCTTCATAGGTTCAGGCTCT	60.7 61.1	152
COMT	MANES_01G043700	F: GGCTGACCACTCAACCATTACC R: GACCGCTCCAGTTCCACCA	60.9 61.2	103
F5H	MANES_04G084300	F: GGTGGCATCGGCAATAGAGTG R: TGCAAGAGGAGTGGGATAGGTG	62.7 61.6	198

Fig. 1 Changes of PAL gene expressions in different varieties at different times before and after mite infestation A: Changes of gene expressions in various varieties before and after mite infestation. B: Changes of gene expressions between varieties at the same time before and after mite infestation. Different lowercase letters indicate that the expression amount after different time of mite infested is significantly different from that before mite infested（P<0.05）. The same below

Fig. 2 Changes of 4 CL gene expressions in different varieties at different times before and after mite infestation

Fig. 3 Changes of C4H gene expressions in different varieties at different times before and after mite infestation

Fig. 4 Changes of HCT gene expressions in different varieties at different times before and after mite infestation

Fig. 5 Changes of CSE gene expressions in different varieties at different times before and after mite infestation

Fig. 6 Changes of COMT gene expressions in different varieties at different times before and after mite infestation

Fig. 7 Changes of CCoAOMT gene expressions in different varieties at different times before and after mite inf-estation

Fig. 8 Changes of F5H gene expressions in different varieties at different times before and after mite infestation

Fig. 9 Changes of CCR gene expressions in different varieties at different times before and after mite infestation

Fig. 10 Changes of CAD gene expressions in different varieties at different times before and after mite infestation

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