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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

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 *** findings were published in a recent issue of Nature Communication[1].Auxin is the plant growth hormone that plays an essential

关键词： auxin crosstalk biosynthesis Xiang hormones Arabidopsis drought signaling abiotic roots

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

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

Synergistic roles of LAX1 and FZP in the development of rice sterile lemma

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The Crop Journal 2020年第1期8卷 16-25页

作者： Yidong Wang Shanshan Wei Yubing He Lang Yan Rongchen Wang yunde zhao aNational Key Laboratory of Crop Genetic Improvement National Center of Plant Gene Research(Wuhan)Huazhong Agricultural UniversityWuhan 430070HubeiChina College of Life Science and Technology Huazhong Agricultural UniversityWuhan 430070HubeiChina Section of Cell and Developmental Biology University of California San DiegoLa JollaCA 92093-0116USA

Rice florets are subtended by two sterile lemmas,whose origin and biological functions have not been studied *** we demonstrate that two putative transcription factors,LAX PANICLE1(LAX1)and FRIZZY PANICLE(FZP),synergistically control the development of sterile *** LAX1 and FZP are previously known for their roles in panicle and floret *** of either LAX1 or FZP greatly reduces the number of floret *** generated new lax1 mutants(lax1-c)using CRISPR/Cas9 gene editing *** addition to the expected lax panicle phenotypes,we noticed that a significant number of spikelets of lax1-c developed elongated sterile ***,our characterization of lax1-RNAi plants also revealed sterile lemma phenotypes similar to lax1-c *** isolated a weak allele of fzp(fzp-14)in a genetic screen for lax1–1 *** fzp-14 lax1–1 double mutants completely eliminated flower ***,the isolated fzp-14 produced spikelets with elongated sterile ***,fzp-14 was haploid-insufficient in the lax1–1 background whereas fzp-14 heterozygous plants were indistinguishable from wild type *** lax1–1 fzp-14+/−also developed elongated sterile lemma as observed in lax1-c,lax1-RNAi,and fzp-14,suggesting that LAX1 and FZP synergistically control sterile lemma development.

关键词： LAX1 FZP Rice CRISPR Flower development Sterile lemma

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

Plant Hormones:Metabolism,Signaling and Crosstalk

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Journal of Integrative Plant Biology 2011年第6期53卷 410-411页

作者： Li-Jia Qu Issue Editor yunde zhao Issue Editor School of Life Sciences Peking University Beijing China Section of Cell and Developmental Biology University of California at San Diego USA

Plants synthesize various hormones in response to environ- mental cues and developmental signals to ensure their proper growth and development. Elucidation of the molecular mech- anisms by which plant hormones control growth and devel- opment contributes to our understanding of fundamental plant biology and provides tools to improve crops. Because of their critical roles in plant growth and development, plant hormones have been studied extensively since the early days of plant biology.

关键词： PLANT hormones PLANT metabolism PLANT cellular signal transduction PLANT development MERISTEMS ABSCISSION (Botany) PLANT growth