Welcome to Acta Agronomica Sinica，

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

RichHTML403

59

PDF

332

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."

[1] Shuai B, Reynagapeña C G, Springer P S. The lateral organ boundaries gene defines a novel, plant-specific gene family. Plant Physiol, 2002, 129: 747-761.
[2] Rubin G, Tohge T, Matsuda F, Saito K, Scheible W. Members of the LBD family of transcription factors repress anthocyanin synthesis and affect additional nitrogen responses in Arabidopsis. Plant Cell, 2009, 21:3567-3584.
doi: 10.1105/tpc.109.067041
[3] Chanderbali A S, He F M, Soltis P S, Soltis D E. Out of the water: origin and diversification of the LBD gene family. Mol Biol Evol, 2015, 32: 1996-2000.
doi: 10.1093/molbev/msv080 pmid: 25839188
[4] Matsumura Y, Iwakawa H, Machida Y. Characterization of genes in the ASYMMETRIC LEAVES2/ LATERAL ORGAN BOUNDARIES (AS2/LOB) family in Arabidopsis thaliana, and functional and molecular comparisons between AS2 and other family members. Plant J, 2009, 58: 525-537.
doi: 10.1111/j.1365-313X.2009.03797.x
[5] Zhang Y M, Zhang S Z, Zheng C C. Genomewide analysis of LATERAL ORGAN BOUNDARIES domain gene family in Zea mays. J Genet, 2014, 93: 79-91.
[6] Okushima Y, Fukaki H, Onoda M, Theologis A, Tasaka M. ARF7 and ARF19 regulate lateral root formation via direct activation of LBD/ASL genes in Arabidopsis. Plant Cell, 2007, 19: 118-130.
pmid: 17259263
[7] Guo B J, Wang J, Lin S, Tian Z, Zhou K, Luan H Y, Lyu C, Zhang X Z, Xu R G. A genome-wide analysis of the ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES (AS2/LOB) gene family in barley (Hordeum vulgare L.). J Zhejiang Univ Sci B, 2016, 17: 763-774.
doi: 10.1631/jzus.B1500277
[8] Inukai Y, Sakamoto T, Ueguchi-Tanaka M, Shibata Y, Gomi K, Umemura I, Hasegawa Y, Ashikari M, Kitano M, Matsuoka M. Crown rootless1, which is essential for crown root formation in rice, is a target of an AUXIN RESPONSE FACTOR in auxin signaling. Plant Cell, 2005, 17: 1387-1396.
doi: 10.1105/tpc.105.030981
[9] Liu H J, Wang S F, Yu X B, Yu J, He X W, Zhang S L, Shou H X, Wu P. ARL1, a LOB-domain protein required for adventitious root formation in rice. Plant J, 2005, 43: 47-56.
doi: 10.1111/j.1365-313X.2005.02434.x
[10] Taramino G, Sauer M, Stauffer J L, Multani D, Niu X M, Sakai H, Hochholdinger F. The maize (Zea mays L.) RTCS gene encodes an LOB domain protein that is a key regulator of embryonic seminal and post-embryonic shoot-borne root initiation. Plant J, 2007, 50: 11.
[11] Ori N, Eshed Y, Chuck G, Bowman J L, Hake S. Mechanisms that control knox gene expression in the Arabidopsis shoot. Development, 2000, 127: 5523-5532.
doi: 10.1242/dev.127.24.5523 pmid: 11076771
[12] Semiarti E, Ueno Y, Tsukay H, Iwakawa H, Machida C, Machida Y. The ASYMMETRIC LEAVES2 gene of Arabidopsis thaliana regulates formation of a symmetric lamina, establishment of venation and repression of meristem-related homeobox genes in leaves. Development, 2001, 128: 1771-1783.
doi: 10.1242/dev.128.10.1771 pmid: 11311158
[13] Byrne M E, Simorowski J, Martienssen R A. ASYMMETRIC LEAVES1 reveals knox gene redundancy in Arabidopsis. Development, 2002, 129: 1957-1965.
doi: 10.1242/dev.129.8.1957 pmid: 11934861
[14] Machida C, Nakagawa A, Kojima S, Takahashi H, Machida Y. The complex of ASYMMETRIC LEAVES (AS) proteins plays a central role in antagonistic interactions of genes for leaf polarity specification in Arabidopsis. Wires Dev Biol, 2015, 4: 655-671.
doi: 10.1002/wdev.196
[15] Oh S A, Park K S, Twell D, Park S K. The SIDECAR POLLEN gene encodes a microspore-specific LOB/AS2 domain protein required for the correct timing and orientation of asymmetric cell division. Plant J, 2010, 64: 839-50.
doi: 10.1111/j.1365-313X.2010.04374.x
[16] Albinsky D, Kusano M, Higuchi M, Hayashi N, Kobayashi M, Fukushima A, Mori M, Ichikawa T, Matsui K, Kuroda H, Horii Y, Tsumoto Y, Sakakibara H, Hirochika H, Matsui M, Saito K. Metabolomic screening applied to rice FOX Arabidopsis lines leads to the identification of a gene-changing nitrogen metabolism. Mol Plant, 2010, 3: 125-142.
doi: 10.1093/mp/ssp069 pmid: 20085895
[17] Chen Y M, Song W J, Xie X M, Wang Z H, Guan P F, Peng H R, Jiao Y N, Ni Z F, Sun Q X, Guo W L. A Collinearity- incorporating homology inference strategy for connecting emerging assemblies in Triticeae tribe as a pilot practice in the plant pangenomic era. Mol Plant, 2020, 13: 1694-1708.
doi: 10.1016/j.molp.2020.09.019
[18] Guo B J, Wei Y F, Xu R B, Lin S, Luan H Y, Lyu C, Zhang X Z, Song X Y, Xu R G. Genome-wide analysis of APETALA2/ ethylene-responsive factor (AP2/ERF) gene family in barley (Hordeum vulgare L.). PLoS One, 2016, 11: e0161322.
[19] Jones H D, Shewry P R, eds. Transgenic wheat, barley and oats:barley transformation using agrobacterium-mediated techniques. In: Methods in Molecular Biology. New York: Humana Press, 2009. pp 137-147.
[20] Majer C, Xu C, Berendzen K W, Hochholdinger F. Molecular interactions of ROOTLESS CONCERNING CROWN AND SEMINAL ROOTS, a LOB domain protein regulating shoot- borne root initiation in maize (Zea mays L.). Philos Trans R Soc Lond B Biol Sci, 2012, 367: 1542-1551.
doi: 10.1098/rstb.2011.0238
[21] Shao Y, Zhou H Z, Wu Y, Zhang H, Lin J, Jiang X Y, He Q J, Zhu J S, Li Y G, Yu H, Mao C Z. OsSPL3, an SBP-domain protein, regulates crown root development in rice. Plant Cell, 2019, 31: 1257-1275.
doi: 10.1105/tpc.19.00038
[22] 张焕欣, 董春娟, 李福凯, 王红飞, 尚庆茂. 植物不定根发生机理的研究进展. 西北植物学报, 2017, 37: 1457-1464.
Zhang H X, Dong C J, Li F K, Wang H F, Shang Q M. Progress on the regulatory mechanism of adventitious rooting. Acta Bot Boreal-Occident Sin, 2017, 37: 1457-1464. (in Chinese with English abstract)
[1] Wang Wen-Yuan, Yan Xue-Jia, Liu Yu-Lin, Sun Xiao-Tong, Li Ya-Nan, Tang Xin-Hua, Shi Ying. Screening of low-light-tolerant potato varieties and cloning and functional analysis of the transcription factor gene StPIF3 [J]. Acta Agronomica Sinica, 2026, 52(6): 1631-1645.
[2] Niu Li, Wang Yong-Sheng, Wang Chang-Jie, Zhang Hong, Meng Ya-Xiong, Li Bao-Chun, Yang Ke, Ma Xiao-Le, Yao Li-Rong, Si Er-Jing, Wang Hua-Jun, Wang Jun-Cheng. Identification and analysis of the NAC gene family in barley (Hordeum vulgare L.) and functional validation of HvNAC38 in salt tolerance [J]. Acta Agronomica Sinica, 2026, 52(3): 688-707.
[3] Li Ying, Shi Xiao-Xu, Liu Hai-Cui, Shi Lyu, Xue Ya-Guang, Wei Ya-Feng. Establishment of a near-infrared reflectance spectroscopy model for predicting β-glucan content in naked barley grain [J]. Acta Agronomica Sinica, 2026, 52(3): 735-745.
[4] Yu Kai-Hang, Zhou Hong-Bin, Luo Liang-Zha, Wang Mei-Li, Jiang Rui-Mei, Dong-Chen Wen-Hua, Li Shi-Jin, Mao Xiao-Qiang, Chen Sheng-Wei. Cloning and expression analysis of the HvLRR-RLK-510 gene encoding a leucine-rich repeat receptor-like kinase in barley [J]. Acta Agronomica Sinica, 2026, 52(2): 421-432.
[5] MA Juan-E, YAO You-Hua, YAO Xiao-Hua, WU Kun-Lun, CUI Yong-Mei. Cloning and functional analysis of HvERF039 gene in Qingke (Hordeum vulgare L. var. nudum Hook. f) [J]. Acta Agronomica Sinica, 2025, 51(9): 2341-2357.
[6] HE Peng-Xu, YAO Li-Rong, CHEN Yuan-Ling, YAN Yan, ZHANG Hong, WANG Jun-Cheng, LI Bao-Chun, YANG Ke, SI Er-Jing, MENG Ya-Xiong, MA Xiao-Le, WANG Hua-Jun. Differences and correlations in physiological and molecular mechanisms of barley germination under drought stress [J]. Acta Agronomica Sinica, 2025, 51(9): 2412-2432.
[7] WEN Xuan, ZHONG Xiu-Li, WANG Shang-Wen, JIN Tao, PENG Jun, LIU En-Ke. Screening of low nitrogen tolerant germplasm in seedling highland barley based on tolerance index and comprehensive evaluation of different nitrogen efficiency types [J]. Acta Agronomica Sinica, 2025, 51(7): 1949-1958.
[8] YIN Yu-Meng, WANG Yan-Nan, KANG Zhi-He, QIAO Shou-Chen, BIAN Qian-Qian, LI Ya-Wei, CAO Guo-Zheng, ZHAO Guo-Rui, XU Dan-Dan, YANG Yu-Feng. Cloning and functional analysis of glutathione S-transferase gene IbGSTU7 in sweetpotato [J]. Acta Agronomica Sinica, 2025, 51(7): 1736-1746.
[9] LU Wen-Jia, WANG Jun-Cheng, YAO Li-Rong, ZHANG Hong, SI Er-Jing, YANG Ke, MENG Ya-Xiong, LI Bao-Chun, MA Xiao-Le, WANG Hua-Jun. Genome-wide identification of PRX gene family and analysis of their expressions under drought stress in barley [J]. Acta Agronomica Sinica, 2025, 51(5): 1198-1214.
[10] CHENG Hong-Na, QIN Dan-Dan, XU Fu-Chao, XU Qing, PENG Yan-Chun, SUN Long-Qing, XU Le, GUO Ying, YANG Xin-Quan, XU De-Ze, DONG Jing. Comparative analysis of metabolomics of colored hulless barley and colored wheat grains [J]. Acta Agronomica Sinica, 2025, 51(4): 932-942.
[11] WANG Lin, CHEN Xiao-Yu, ZHANG Wen-Meng-Long, WANG Si-Qi, CHENG Bing-Yun, CHENG Jing-Qiu, PAN Rui, ZHANG Wen-Ying. Molecular characteristics and functional analysis of HvMYB2 in response to drought stress in barley [J]. Acta Agronomica Sinica, 2025, 51(4): 873-887.
[12] ZHANG Hui-Min, DANG Ya-Ru, SANG Jia-Nan, JIAO Wen-Jing, SUN Chun-Yi, ZHUANG Jia-Mu, WEI Yu-Shu, ZHANG Chao, TIAN Peng, LIU Bai-Lin, SONG Yin. Genome-wide identification of CEP gene family in potato (Solanum tuberosum L.) and functional analysis of StCEP2 in regulating adventitious root growth and salt tolerance [J]. Acta Agronomica Sinica, 2025, 51(12): 3224-3237.
[13] LIU Jia-Hui, LI Yu-Long, WANG Ya-Ru, HE Hong, ZHANG Yun-Shu, WU Yu, ZENG Xiu-Li, LIU Ting-Hui, CHEN Guo-Yue, QI Peng-Fei, WEI Yu-Ming, JIANG Qian-Tao. Effects of natural variation in the SSIIa gene on starch composition and properties in Tibetan barley [J]. Acta Agronomica Sinica, 2025, 51(12): 3144-3156.
[14] MA Min-Hu, CHANG Hua-Yu, CHEN Zhao-Yan, REN Zeng, LIU Ting-Hui, XING Guo-Fang, GUO Gang-Gang. Identification and genome-wide association study of specialized seedling grass barley cultivars [J]. Acta Agronomica Sinica, 2025, 51(1): 91-102.
[15] SUN Man, AN Chao-Dan, GAO Guang-Qi, GUO Jie, YANG Ping, JIANG Cong-Cong. Genetic dissection of the albino hull mutations in barley (Hordeum vulgare L.) [J]. Acta Agronomica Sinica, 2024, 50(12): 3046-3054.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd