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Epigenetic Suppression of T-DNA Insertion Mutants in Arabidopsis

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Molecular Plant 2013年第2期6卷 539-545页

作者： Yangbin Gao yunde zhao Section of Cell and Developmental Biology University of California San Diego 9500 Gilman Drive La Jolla CA 92093-0116 USA

T-DNA insertion mutants have been widely used to define gene functions in Arabidopsis and in other plants. Here, we report an unexpected phenomenon of epigenetic suppression of T-DNA insertion mutants in Arabidopsis. When the two T-DNA insertion mutants, yucl-1 and ag-TD, were crossed together, the defects in all of the ag-TD plants in the F2 popumation were partially suppressed regardless of the presence of yucl-1. Conversion of ag-TD to the suppressed ag- TD （named as ag-TD＊） did not follow the laws of Mendelian genetics. The ag-TD＊ could be stably transmitted for many generations without reverting to ag-TD, and ag-TD^＊ had the capacity to convert ag-TD to ag-TD^＊. We show that epige-netic suppression of T-DNA mutants is not a rare event, but certain structural features in the T-DNA mutants are needed in order for the suppression to take place. The suppressed T-DNA mutants we observed were all intronic T-DNA mutants and the T-DNA fragments in both the trigger T-DNA as well as in the suppressed T-DNA shared stretches of identical sequences. We demonstrate that the suppression of intronic T-DNA mutants is mediated by trans-interactions between two T-DNA insertions. This work shows that caution is needed when intronic T-DNA mutants are used.

关键词： epigenetics T-DNA mutant genetic suppression trans-interaction YUC.

A Role for Auxin in Flower Development

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Journal of Integrative Plant Biology 2007年第1期49卷 99-104页

作者： Youfa Cheng yunde zhao Section of Cell and Developmental Biology University of California San Diego La Jolla CA 92093-0116 USA

Auxin has long been implicated in many aspects of plant growth and development including flower development. However, the exact roles of auxin in flower development have not been well defined until the recent identification of auxin biosynthesis mutants. Auxin is necessary for the inltiation of floral primordia, and the disruption of auxin biosynthesis, polar auxin transport or auxin signaling leads to the failure of flower formation. Auxin also plays an essential role in specifying the number and Identity of floral organs. Further analysis of the relationship between the auxin pathways and the known flower development genes will provide critical information regarding mechanisms of organogenesis and pattern formation in plants.

关键词： auxin development flower

Auxin Biosynthesis： A Simple Two-Step Pathway Converts Tryptophan to Indole-3-Acetic Acid in Plants

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Molecular Plant 2012年第2期5卷 334-338页

作者： yunde zhao Section of Cell and Developmental Biology University of California San Diego 9500 Gilman Drive La Jolla CA 92093-0116 USA

Indole-3-acetic acid （IAA）, the main naturally occurring auxin, is essential for almost every aspect of plant growth and development. However, only recently have studies finally established the first complete auxin biosynthesis pathway that converts tryptophan （Trp） to IAA in plants. Trp is first converted to indole-3-pyruvate （IPA） by the TAA family of amino transferases and subsequently IAA is produced from IPA by the YUC family of flavin monooxygenases. The two- step conversion of Trp to IAA is the main auxin biosynthesis pathway that plays an essential role in many developmental processes.

关键词： hormonal regulation hormone biology genetics development auxin.

Technological breakthroughs in generating transgene-free and genetically stable CRISPR-edited plants

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aBIOTECH 2020年第1期1卷 88-96页

作者： Yubing He yunde zhao National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research(Wuhan) Huazhong Agricultural UniversityWuhan 430070China Section of Cell and Developmental Biology University of California San DiegoLa JollaCA 92093-0116USA

CRISPR/Cas9 gene-editing technologies have been very effective in editing target genes in all major crop plants and offer unprecedented potentials in crop improvement.A major challenge in using CRISPR gene-editing technology for agricultural applications is that the target gene-edited crop plants need to be transgene free to maintain trait stability and to gain regulatory approval for commercial *** this article,we present various strategies for generating transgene-free and target geneedited crop *** CRISPR transgenes can be removed by genetic segregation if the crop plants are reproduced ***-assisted tracking and eliminating transgenes greatly decrease the time and labor needed for identifying the ideal transgene-free *** can be programed to undergo self-elimination when CRISPR genes and suicide genes are sequentially activated,greatly accelerating the isolation of transgene-free and target gene-edited ***-free plants can also be generated using approaches that are considered non-transgenic such as ribonucleoprotein transfection,transient expression of transgenes without DNA integration,and ***,we discuss the advantages and disadvantages of the various strategies in generating transgene-free plants and provide guidance for adopting the best strategies in editing a crop plant.

关键词： CRISPR Transgene-free Marker-assisted selection TKC Cas9 gene editing

A key link between jasmonic acid signaling and auxin biosynthesis

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Science China(Life Sciences) 2015年第3期58卷 311-312页

作者： HE YuBing zhao yunde National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research Huazhong Agricultural University Section of Cell and Developmental Biology University of California San Diego

A research team led by Xiang Cheng Bin from School of Life Sciences,University of Science and Technology of China discovered a molecular basis for crosstalk between jasmonic acid(JA)and auxin,two important plant *** f... 详细信息

关键词：生长素茉莉酸生物合成中国科学技术大学信号生命科学学院植物生长激素分子基础

Revolutionize Genetic Studies and Crop Improvement with High-Throughput and Genome-Scale CRISPR/Cas9 Gene Editing Technology

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Molecular Plant 2017年第9期10卷 1141-1143页

作者： Yang, Ning Wang, Rongchen zhao, yunde Huazhong Agr Univ Natl Key Lab Crop Genet Improvement Wuhan 430070 Hubei Peoples R China Huazhong Agr Univ Natl Ctr Plant Gene Res Wuhan Wuhan 430070 Hubei Peoples R China Univ Calif San Diego Sect Cell & Dev Biol La Jolla CA 92093 USA

Forward genetic screens have been instrumental in deciphering the molecular mechanisms that control agriculturally important traits. Traditionally, crop mutant collections are generated by chemical mutagenesis such as ethyl methanesulfonate （EMS） treatment, physical irradiation, and insertional agents including T-DNAs and transposons （Wei et al., 2013; Li et al., 2016）.

关键词： Revolutionize Genetic Studies Crop Improvement High-Throughput

NPY Genes Play an Essential Role in Root Gravitropic Responses in Arabidopsis

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Molecular Plant 2011年第1期4卷 171-179页

作者： Yuanting Li Xinhua Dai Youfa Cheng yunde zhao Section of Cell and Developmental Biology University of California San Diego 9500 Gilman Drive La Jolla CA92093-0116 USA Key Laboratory of Arid and Grassland Agroecology of Ministry of Education School of Life Sciences Lanzhou University Lanzhou 730000 China Present address: Key Laboratory of Photosynthesis and Environmental Molecular Physiology Institute of Botany Chinese Academy of Sciences Beijing 100093 China

Plants can sense the direction of gravity and orient their growth to ensure that roots are anchored in soil and that shoots grow upward. Gravitropism has been studied extensively using Arabidopsis genetics, but the exact mecha- nisms for gravitropism are not fully understood. Here, we demonstrate that five NPY genes play a key role in Arabidopsis root gravitropism. NPYgenes were previously identified as regulators of auxin-mediated organogenesis in a genetic pathway with the AGC kinases PID, PID2, WAG1, and WAG2. We show that all five NPYgenes are highly expressed in primary root tips. The single npy mutants do not display obvious gravitropism defects, but the npyl npy2 npy3 npy4 npy5 quin- tuple mutants show dramatic gravitropic phenotypes. Systematic analysis of all the npy double, triple, and quadruple combinations demonstrates that the five NPY genes all contribute to gravitropism. Our work indicates that gravitropism, phototropism, and organogenesis use analogous mechanisms in which at least one AGC kinase, one NPH3/NPY gene, and one ARF are required.

关键词： Light signaling signal transduction development root biology auxin.

ESCRT-dependent vacuolar sorting and degradation of the auxin biosynthetic enzyme YUC1 flavin monooxygenase

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Journal of Integrative Plant Biology 2019年第9期61卷 968-973页

作者： Chennan Ge Caiji Gao Qingguo Chen Liwen Jiang yunde zhao National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research(Wuhan) Huazhong Agricultural UniversityWuhan430070China Section of Cell and Developmental Biology University of CaliforniaSan DiegoLa JollaCA92093-0116USA School of Life Sciences The Chinese University of Hong KongShatinNew TerritoriesHong KongChina Guangdong Provincial Key Laboratory of Biotechnology for Plant Development School of Life SciencesSouth China Normal UniversityGuangzhou510631China

YUC flavin monooxygenases catalyze the ratelimiting step of auxin biosynthesis. Here we report the vacuolar targeting and degradation of GFP-YUC1. GFP-YUC1 fusion expressed in Arabidopsis protoplasts or transgenic plants was primarily localized in vacuoles. Surprisingly, we established that GFP-YUC1, a soluble protein, was sorted to vacuoles through the ESCRT pathway, which has long been recognized for sorting and targeting integral membrane proteins. We further show that GFP-YUC1 was ubiquitinated and in this form GFP-YUC1 was targeted for degradation, a process that was also stimulated by elevated auxin levels. Our findings revealed a molecular mechanism of GFP-YUC1 degradation and demonstrate that the ESCRT pathway can recognize both soluble and integral membrane proteins as cargoes.

关键词： GFP PVC bar TAA ESCRT-dependent vacuolar

A reporter for noninvasively monitoring gene expression and plant transformation

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Horticulture Research 2020年第1期7卷 821-826页

作者： Yubing He Tao Zhang Hui Sun Huadong Zhan yunde zhao State Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural UniversityNanjing 210095China National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research(Wuhan) Huazhong Agricultural UniversityWuhan 430070China Section of Cell and Developmental Biology University of California San Diego9500 Gilman DriveLa JollaCA 92093-0116USA Tata Institute for Genetics and Society-UCSD La JollaCA 92093-0335USA

Reporters have been widely used to visualize gene expression,protein localization,and other cellular activities,but the commonly used reporters require special equipment,expensive chemicals,or invasive ***,we construct a new reporter RUBY that converts tyrosine to vividly red betalain,which is clearly visible to naked eyes without the need of using special equipment or chemical *** show that RUBY can be used to noninvasively monitor gene expression in ***,we show that RUBY is an effective selection marker for transformation events in both rice and *** new reporter will be especially useful for monitoring cellular activities in large crop plants such as a fruit tree under field conditions and for observing transformation and gene expression in tissue culture under sterile conditions.

关键词： transformation reporter sterile