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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (1): 134-148.doi: 10.3724/SP.J.1006.2025.44035

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Analysis of expression patterns of laccase gene family members in Brassica napus and their association with stem fracture resistance

XU Lin-Shan1(), GAO Geng-Dong1, WANG Yu1, WANG Jia-Xing1, YANG Ji-Zhao1, WU Ya-Rui1, ZHANG Xiao-Han1, CHANG Ying1, LI Zhen1, XIE Xiong-Ze2, GONG De-Ping3, WANG Jing1,*(), GE Xian-Hong1   

  1. 1College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, China
    2Xiangyang Academy of Agricultural Sciences, Xiangyang 441057, Hubei, China
    3Jingzhou Academy of Agricultural Sciences, Jingzhou 434000, Hubei, China
  • Received:2024-02-27 Accepted:2024-08-15 Online:2025-01-12 Published:2024-09-02
  • Contact: *E-mail: wangjing@mail.hzau.edu.cn
  • Supported by:
    National Key Research and Development Program ‘Creation and Application of New Germplasm of Multi-resistance Rapeseed’(2023YFD1201402);Hubei Key Research and Development Program ‘High-throughput Intelligent Selection and Breeding of High-yield Rapeseed Varieties with High Resistance to Dense Planting’(2023BBB030)

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Abstract:

Laccase is a family of copper-containing polyphenol oxidases primarily involved in lignin synthesis and resistance to various stresses in plants. In this study, members of the laccase gene family (BnaLACs) in Brassica napus were identified, and their physical and chemical properties were measured, including the number of amino acids, molecular weight, isoelectric point, instability index, and aliphatic index. The chromosome positions, evolutionary relationships, gene structures, and tissue expression patterns of these genes were subsequently predicted and analyzed. The results showed that the Brassica napus genome contains 53 BnaLAC family members, which are generally alkaline and stable proteins. Most BnaLACs are located in the vacuole membrane and outside the cell. Gene structure analysis revealed that BnaLACs have conserved structures. Tissue expression pattern analysis indicated that BnaLACs are expressed in all tissues except anthers, with higher expression levels in roots, seeds, silique walls, and stems. The expression pattern of BnaLAC4s in stems was specifically analyzed, and it was found that BnaA05G0074200ZS is significantly correlated with lodging resistance in Brassica napus. Haplotype analysis showed significant differences in lodging resistance and lignin content between different BnaA05G0074200ZS haplotype lines. The results of this study provide a foundation for further analysis of the functions of the laccase gene family in Brassica napus and the mechanisms underlying stem lodging resistance.

Key words: Brassica napus, laccase, evolution, expression pattern, haplotype, lodging resistance

Table 1

Primers used for BnaA05G0074200ZS validation"

引物类型Primer type 引物名称Primer name 引物序列Primer sequence (5°-3°)
单倍型鉴定引物
Haplotype identification primer		 BnaLAC-A5-3329-F1 CTACTGTTGTTCGTCGTTTCTATTG
BnaLAC-A5-3329-R2 CTACTCACACACATTATTGTGGCT
BnaA5LAC-3K-F2 CTCCTCGGTTCTCTTTCTTCTA
BnaA5LAC-3K-R2 CATCTAATTGTATGTCTTATCGCCTTT
qRT-PCR引物
qRT-PCR primer		 qRTBnaLAC-A5F2 AGGCGCTTAAGTCTGGATTAGCCC
qRTBnaLAC-A5R2 AACGGTTTGACGTAGACAGCGTCC
BnENTH-F GTTTAGACCCGTTGCTGCTC
BnENTH-R TTGTCCATCTCAGCCATTTG

Fig. 1

Distribution of BnaLACs on chromosomes in Brassica napus"

Fig. 2

Phylogenetic tree of LAC family members in Arabidopsis thaliana and Brassica napus There are nine categories, each marked with a different color."

Fig. 3

Analysis of conserved motifs and gene structures of LAC family members in Arabidopsis thaliana and Brassica napus According to the phylogenetic relationship, the BnaLAC family was divided into nine categories, each of which was marked with a different color. A total of ten conserved motifs have been identified by the BnaLAC family, with different motifs represented by squares of different colors. Yellow squares in the gene structure represent exons, green squares represent UTR regions, and black lines represent introns."

Fig. 4

Tissue expression patterns of BnaLACs family throughout the growth period of Brassica napus The expression of BnaLACs in each tissue were 2 mm and 4 mm buds, filament, petal, pollen, sepal, cotyledon, vegetative rosette, 1-23 leaves, root, seeds 14-64 days after flowering, silique wall 2- -60 days after flowering, middle, and upper stems."

Fig. 5

Expression patterns of BnaLACs family members in different tissue A: BnaLACs expression in cotyledon, vegetative rosette and 1-23 leaves. B: BnaLACs expression in seeds 14-64 days after flowering. C: expression level of BnaLACs in silique wall 2-60 days after flowering. D: BnaLACs expression in lower, middle and upper stems. E: expression level of BnaLACs in root. F: expression of BnaLACs in buds of 2 mm and 4 mm. G: expression of BnaLACs in filament."

Fig. 6

BnaLAC4s expression pattern and lignin content in ZS11 and lodging cultivar L23 A, B, and C are the expression patterns of ZS11 in Brassica napus database. D, E, and F are the expression patterns of BnaLAC4s in ZS11 and L23, ZS11: Zhongshuang 11. G: lignin content. Student’s t-test, * indicates significant difference at the 0.05 probability level."

Fig. 7

Haplotype distribution of BnaA05G0074200ZS in natural populations A: haplotype type of BnaA05G0074200ZS. B: relationship between BnaA05G0074200ZS haplotype and flexural strength in natural population; Student's t test, P=0.016 < 0.05, the difference was significant. C: Relative expression levels of BnaA05G0074200ZS in different haplotypes, hap1 for C152 and C344, hap2 for C275 and C474. Different letters indicate significant difference at the P < 0.05 level by one-way ANOVA. D: Lignin content for different haplotypes."

Table 2

BnaLAC family members and physicochemical properties"

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