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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (10): 2435-2442.doi: 10.3724/SP.J.1006.2022.11088

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

Regulation of adventitious root development by HvLBD19 gene in barley (Hordeum vulgare L.)

GUO Bao-Jian1,2(), WANG Shuang1,2, LYU Chao1,2, WANG Fei-Fei1,2, XU Ru-Gen1,2,*()   

  1. 1Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding / Key Laboratory of Plant Functional Genomics of the Ministry of Education / Jiangsu Key Laboratory of Crop Genetics and Physiology, Agricultural College of Yangzhou University, Yangzhou 225009, Jiangsu, China
    2Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, Jiangsu, China
  • Received:2021-10-12 Accepted:2022-02-25 Online:2022-10-12 Published:2022-03-22
  • Contact: XU Ru-Gen E-mail:bjguo@yzu.edu.cn;rgxu@yzu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(31771771);Natural Science Foundation of the Jiangsu Higher Education Institutions of China(19KJA560005);China Agriculture Research System of MOF and MARA(CARS-05);and the Priority Academic Program Development of Jiangsu Higher Education Institutions

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Abstract:

LBD gene family is a plant specific transcription factor family, which plays an important role in regulating plant development and nitrogen metabolism. Micro-synteny analysis showed that barley HvLBD19 was an orthologous of maize RTCS and rice ARL1 gene. Temporal and spatial expression analysis revealed that this gene was most abundant in adventitious roots and was induced by exogenous auxin treatment, which encoded a protein located in the nucleus. Transgenic results indicated that the adventitious root length increased by nearly one time, and the number of adventitious roots increased by 40%. It was preliminarily confirmed that HvLBD19 gene controlled adventitious root development in barley and provided a foundation for further research on the molecular mechanism of HvLBD19 gene in regulating adventitious root development in barley.

Key words: barley, LATERAL ORGAN BOUNDARIES DOMAIN gene, overexpression, adventitious root

Table 1

Primers used in the present study"

引物编号
Primer pairs ID		 引物序列
Primer sequence (5′-3′)		 内容
Content
P1 ATGACGGGACTGGGTTCGCC 编码区扩增
Coding sequence amplification
TTACGAGCGATTAAGGTAAG
P2 AGTTTCTGCGTCGCAAGTG 荧光定量
qRT-PCR
GAGCTTGGAGACGTTGCTG
P3 GGTCCATCCTAGCCTCACTC 内参基因HvActin扩增
Internal control of HvActin amplification
GATAACAGCAGTGGAGCGCT
P4 TCTCCCTTACCCATGATGAGCATGACGGGACTGGG 瞬时表达载体构建
Transient expression vector construction
CACGGGGGACTCTAGCGAGCGATTAAGGTAAGCATACGC
P5 CCCAAGCTTCTTGCATGCCTGCAGTGCAGCGTGACCCG Ubi启动子扩增
Ubi promoter amplification
CCCCCCGGGCTGCAGAAGTAACACCAAAC
P6 GACTAGTGAGCTCGAATTTCCCCGATC nos终止子扩增
nos terminator amplification
GCTCTAGAGATCTGATATCATCGATGAA
P7 CCCCCCGGGATGACGGGACTGGGTTCGCC 大麦超表达载体构建
Overexpression vector construction in barley
GACTAGTTTACGAGCGATTAAGGTAAG

Fig. 1

Construction of HvLBD19 gene overexpression vectors in barley A: the construction of tobacco transient expression vector; B: the construction of barley overexpression vector."

Fig. 2

Genetic transformation of barley A: young spike; B: callus induction of immature embryos; C: callus transformation; D: regenerating transformed barley line."

Fig. 3

Sequence analysis of LBD genes A: sequence alignment of HvLBD19, OsARL1, and ZmRTCS proteins; B: micro-synteny analysis of the LBD gene in maize, barley, and rice."

Fig. 4

Relative expression profiling of HvLBD19 gene A: leaf (L), adventitious root (AR), node root (NR), and seminal root (SR) (gene expression was represented as the relative expression to that for leaf); B and C: the relative expression profiling in seminal root and node root under NAA treatment (Gene expression was represented as the relative expression to that for 0 h)."

Fig. 5

Subcellular location of HvLBD19 protein Bar: 20 μm."

Fig. 6

Genetic transformation and positive plants identification in barley A: PCR identification of positive plants; P: plasmid; G: Golden Promise; numbers: transgenic lines; M: DNA marker. B: hygromycin screening. C: the relative expression level of HvLBD19 gene between Golden Promise and transgenic lines. * and ** indicate the significant difference at the 0.01 and 0.05 probalitity levels between transgenic lines and wild type, respectively."

Fig. 7

Observation on root traits of seedling stage A: phenotype of wild-type and transgenic lines; B: seminal root length; C: number of seminal roots; D: adventitious root length; E: number of adventitious roots."

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