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CRISPR/Cas9-mediated disruption of TaNP1 genes results in complete male sterility in bread wheat

Journal of Genetics and Genomics 2020年第5期47卷 263-272页

作者： Jian Li Zheng Wang Guangming He Ligeng Ma Xing Wang Deng Peking University Institute of Advanced Agricultural Sciences WeifangShandong261325China State Key Laboratory of Protein and Plant Gene Research Peking-Tsinghua Center for Life SciencesSchool of Advanced Agriculture Sciences and School of Life SciencesPeking UniversityBeijing100871China College of Life Sciences Capital Normal UniversityBeijing Key Laboratory of Plant Gene Resources and Biotechnology for Carbon Reduction and Environmental ImprovementBeijing100048China

male sterile genes and mutants are valuable resources in hybrid seed production for monoclinous *** genetic redundancy due to allohexaploidy makes it difficult to obtain the nuclear recessive male sterile mutants through spontaneous mutation or chemical or physical mutagenesis methods in *** emerging effective genome editing tool,CRISPR/Cas9 system,makes it possible to achieve simultaneous mutagenesis in multiple *** improve the genome modification efficiency of the CRISPR/Cas9 system in wheat,we compared four different RNA polymerase(Pol)Ⅲpromoters(TaU3 p,TaU6 p,OsU3 p,and OsU6 p)and three types of sgRNA scaffold in the protoplast *** show that the TaU3 promoter-driven optimized sgRNA scaffold was most *** optimized CRISPR/Cas9 system was used to edit three TaNP1 homoeoalleles,whose orthologs,OsNP1 in rice and ZmIPE1 in maize,encode a putative glucose-methanol-choline oxidoreductase and are required for male *** homozygous mutations in TaNP1 genes result in complete male *** further demonstrated that anyone wild-type copy of the three TaNP1 genes is sufficient for maintenance of male *** together,this study provides an optimized CRISPR/Cas9 vector for wheat genome editing and a complete male sterile mutant for development of a commercially viable hybrid wheat seed production system.

关键词： Wheat CRISPR/Cas9 TaNP1 genes male sterility

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Characterization and Use of male sterility in Hybrid Rice Breeding

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Journal of Integrative Plant Biology 2007年第6期49卷 791-804页

作者： Shaoqing Li Daichang Yang Yingguo Zhu Key Laboratory of MOE for Plant Developmental Biology and Institute of Genetics College of Life Sciences Wuhan University Wuhan 430072 China

The hybrid rice （Oryza sativa L.） breeding that was Initiated In China in the 1970s led to a great improvement in rice productivity. In general, It increases the grain yield by over 20% to the inbred rice varieties, and now hybrid rice has been widely introduced into Africa, Southern Asia and America. These hybrid varieties are generated through either three-line hybrid and two-line hybrid systems; the former is derived from cytoplasmic male sterility （CMS） and the latter derived from genlc male sterility （GMS）. There are three major types of CMS （HL, BT and WA） and two types of GMS （photoperlod-sensitlve （PGMS） and temperature-sensitive （TGMS））. The BT- and HL-type CMS genes are characterized as orf79 and orfH79, which are chimeric toxic genes derived from mltochondrial rearrangement. Rf3 for CMS-WA Is located on chromosome 1, while Rf1, Rf4, Rf5 and Rf6 correspond to CMS-BT, CMSoWA and CMS- HL, located on chromosome 10. The Rfl gene for BT-CMS has been cloned recently, and encodes a mltochondriatargeted PPR protein. PGMS Is thought to be controlled by two recessive loci on chromosomes 7 and 12, whereas nine recessive alleles have been identified for TGMS and mapped on different chromosomes. Attention Is still urgently needed to resolve the molecular complexity of male sterility to assist rice breeding.

关键词： hybrid rice male sterility Oryza sativa.

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Jasmonic acid-mediated stress responses share the molecular mechanism underlying male sterility induced by deficiency of ZmMs33 in maize

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The Crop Journal 2023年第4期11卷 1115-1127页

作者： Ziwen Li Shuangshuang Liu Taotao Zhu Jing Wang Meng Sun Xueli An Xun Wei Cuimei Liu Jinfang Chu Xiangyuan Wan Research Institute of Biology and Agriculture Shunde Innovation SchoolUniversity of Science and Technology BeijingBeijing 100024China Zhongzhi International Institute of Agricultural Biosciences Beijing 100192China Beijing Engineering Laboratory of Main Crop Bio-Tech Breeding Beijing International Science and Technology Cooperation Base of Bio-Tech BreedingBeijing Solidwill Sci-Tech Co.Ltd.Beijing 100192China National Centre for Plant Gene Research(Beijing) Chinese Academy of SciencesBeijing 100101China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing 100039China

Plant male reproduction is a fine-tuned developmental process that is susceptible to stressful environments and influences crop grain *** signaling functions in control of plant normal growth and development as well as in response to external stresses,but the interaction or crosstalk among phytohormone signaling,stress response,and male reproduction in plants remains poorly ***-species comparison among 514 stress-response transcriptomic libraries revealed that ms33-6038,a genic male sterile mutant deficient in the Zm Ms33/Zm GPAT6 gene,displayed an excessive drought stress-like transcriptional reprogramming in anthers triggered mainly by disturbed jasmonic acid(JA)*** increased level of JA appeared in Zm Ms33-deficient anthers at both meiotic and postmeiotic stages and activated genes involved in JA biosynthesis and signaling as well as genes functioning in JA-mediated drought *** accumulation of JA elevated expression level of a gene encoding a WRKY transcription factor that activated the Zm Ms33 *** findings reveal a feedback loop of Zm Ms33-JA-WRKY-Zm Ms33 in controlling male sterility and JA-mediated stress response in maize,shedding light on the crosstalk of stress response and male sterility mediated by phytohormone homeostasis and signaling.

关键词： ZmMs33/ZmGPAT6 Jasmonic acid Phytohormone homeostasis male sterility Stress response

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An epi-allele of SMS causes Sanming dominant genic male sterility in rice

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Science China(Life Sciences) 2023年第12期66卷 2701-2710页

作者： Yachun Yang Cheng Zhang Hao Li Zeyuan Yang Zuntao Xu Dewei Tai Dahu Ni Pengcheng Wei Chengxin Yi Jianbo Yang Yong Ding Rice Research Institute Anhui Academy of Agricultural SciencesHefei 230036China Institute of Health and Medicine Hefei Comprehesive National Science CenterHefei 230031China Ministry of Education Key Laboratory for Membraneless Organelles and Cellular Dynamics School of Life SciencesDivision of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefei 230027China

male sterility is an important trait in rice for hybrid rice(Oryza sativa)***,the factors involved in dominant male sterility are largely ***,we identified a gene from Sanming dominant genic male sterile rice,named Sanming dominant male sterility(SMS),and reported that an epi-allele of this locus contributes to male *** analysis attributed dominant male sterility to a single locus,SMS,which we characterized using a male-sterile near isogenic line(NIL)of rice cultivar *** SMS locus was heterozygous in the male-sterile 93-11 NIL,containing an epi-allele identical to that in 93-11,and an epi-allele identical to that in rice cultivar Nipponbare,which we refer to as SMS9 and SMSN,***9is silent and hyper-methylated,whereas SMSNis expressed and hypo-methylated in the 93-11 *** SMSNled to male *** in SMS rescued the male sterility of the 93-11 ***,we observed the duplication of SMSN in Nipponbare,but did not observe the duplication of SMS9in ***,these findings suggest that the reduced methylation and enhanced expression of the SMSNepi-allele in the 93-11 NIL is responsible for its role in conferring dominant male sterility.

关键词： male sterility rice epi-allele Sanming DNA methylation

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A Fragment Substitution in Promoter of MS92/PTC1 Causes male sterility in Rice

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Rice science 2020年第5期27卷 396-404,I0018-I0023页

作者： Qin Peng Deng Luchang Chen Weilan Huang Juan Fan Shijun Tu Bin Tan Jun Yuan Hua Wang Yuping Ma Bingtian Li Shigui Rice Research Institute/State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China Sichuan Agricultural UniversityChengdu 611130China Crop Research Institute Sichuan Academy of Agricultural SciencesChengdu 610066China Characteristic Crop Research Institute Chongqing Academy of Agricultural SciencesChongqing 402160China

Persistent tapetal cell1(PTC1) plays a curial role in pollen development, and is thought to function as a transcriptional activator in rice. However, the molecular mechanism of PTC1 in regulating pollen development and its cis-elements are not well understood. We identified a novel weak male sterility mutant(ms92) which exhibited expanded tapetum and shrink pollen grains. Map-based cloning and allelic analysis suggested that the male sterility of ms92 was caused by a DNA fragment substitution in the promoter of PTC1. The decreased expression of MS92/PTC1 in ms92 and cis-element analysis indicated that the substituted sequence contained several potential binding cis-element of negative feedback. MS92/PTC1 was specifically expressed in tapetum and microspores at the young microspore stage, and its protein was localized in nucleus. We further found that MS92/PTC1 functions as a transcription activator by recognizing H3K4me3. Transcriptomic analysis revealed that a number of genes involved in tapetum degeneration and pollen wall formation were down-regulated in ms92, which are the potential targets of MS92/PTC1. The substitution fragment in MS92/PTC1 promoter was essential for pollen development, and we provided a novel mutant for further identifying the cis-elements in promoter and the molecular network of MS92/PTC1.

关键词： rice MS92/PTC1 plant homeodomain finger cis-element male sterility anther

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Plant male sterility Induced by Anti-Gene CYP86MF in Brassica oleracea var. italica

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Agricultural Sciences in China 2005年第11期4卷 806-810页

作者： HUANG Ke CAO Jia-shu YU Xiao-lin YE Wan-zhi LU Gang XIANG Xun Institute of Vegetable Science Zhejiang University Hangzhou 310029 P.R.China Research Center of Vegetable Science Fujian Academy of Agricultural Sciences Fuzhou 350013 P.R.China

An anti-gene CYP86MF was introduced into hypocotyls of broccoli （Brassica oleracea ***. italica Plenck） with Agrobacterium tumefaciens, and the transgenic plants were obtained by kanamycin selection. The results of PCR, Southern blot and Northern blot indicated that the anti-CYP86MF has been integrated into chromosome of the transgenic plant. And also, plants with hypogenetic stamina or ungerminated pollen were observed. The transgenic male sterility plant could fructify via artificial pollination with normal pollen. Thus it was proved that the pistil of male sterility plant was normally developed, and the sterility originated from anti-CYP86MF.

关键词： Brassica oleracea Broccoli Anti-gene male sterility

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Characterization and mapping of a new male sterility mutant of anther advanced dehiscence(t) in rice

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Journal of Genetics and Genomics 2008年第3期35卷 177-182页

作者： Yi Zhang Yunfeng Li Jian Zhang Fucheng Shen Yuanxin Huang Zhiwei Wu Key Laboratory of Biotechnology and Crop Quality Improvement Ministry of Agriculture Southwest University Chongqing 400716 China

Anther dehiscence is very important for pollen maturation and release. The mutants of anther dehiscence in rice （Oryza sativa L.） are few, and related research remains poor. A male sterility mutant of anther dehiscence in advance, add（t）, has been found in Minghui 63 and its sterility is not sensitive to thermo-photo. To learn the character of sterilization and the function of the add（t） gene, the morphological and cytological studies on the anther and pollen, the ability of the pistil being fertilized, inheritance of the mutant, and mapping of add（t） gene have been conducted. The anther size is normal but the color is white in the mutant against the natural yellow in the wild-type. The pollen is malformed, unstained, and small in the KI-I2 solution. The anther dehiscence is in advance at the bicellular pollen stage. A crossing test indicated that the grain setting ratio of the add（t） is significantly lower than that of the CMS line 2085A. The ability of the pistil being fertilized is most probably decreased by the add（t） gene. The male sterility is controlled by a single recessive gene of add（t）. This gene is mapped between the markers of R02004 （InDel） and RM300 （SSR） on chromosome 2, and the genetic distance from the add（t） gene to these markers is 0.78 cM and 4.66 cM, respectively.

关键词： male sterility sterility character gene mapping rice

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Tribenuron-Methyl Induces male sterility through Anther-Specific Inhibition of Acetolactate Synthase Leading to Autophagic Cell Death

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Molecular Plant 2015年第12期8卷 1710-1724页

作者： Lun Zhao Xue Jing Li Chen Yingjun Liu Yanan Su Tingting Liu Changbin Gao Bin Yi Jing Wen Chaozhi Ma Jinxing Tu Jitao Zou Tingdong Fu Jinxiong Shen National Key Laboratory of Crop Genetic Improvement National Center of Rapeseed Improvement Huazhong Agricultural University Wuhan 430070 China National Research Council Canada Saskatoon Saskatchewan S7N 0W9 Canada

Tribenuron-methyl （TM） is a powerful sulfonylurea herbicide that inhibits branched-chain amino acid （BCAA） biosynthesis by targeting the catalytic subunit （CSR1） of acetolactate synthase （ALS）. Selective in- duction of male sterility by foliar spraying of TM at low doses has been widely used for hybrid seed produc- tion in rapeseed （Brassica napus）; however, the underlying mechanism remains unknown. Here, we report greater TM accumulation and subsequent stronger ALS inhibition and BCAA starvation in anthers than in leaves and stems after TM application. Constitutive or anther-specific expression of csrl-lD （a CSR1 mutant） eliminated anther-selective ALS inhibition and reversed the TM-induced male sterile phenotype in both rapeseed and Arabidopsis. The results of TM daub-stem experiments, combined with the observations of little TM accumulation in anthers and reversion of TM-induced male sterility by targeted expression of the TM metabolism gene Bel in either the mesophyll or phloem, suggested that foliar-sprayed TM was polar-transported to anthers mainly through the mesophyll and phloem. Microscopy and immunoblotting revealed that autophagy, a bulk degradation process induced during cell death, was elevated in TM-induced male sterile anthers and by anther-specific knockdown of ALS. Moreover, TM-induced pollen abortion was significantly inhibited by the autophagy inhibitor 3-MA. These data suggested that TM was polar-transported to anthers, resulting in BCAA starvation via anther-specific ALS inhibition and, ulti- mately, autophagic cell death in anthers.

关键词： male sterility chemical hybridizing agent acetolactate synthase amino acid starvation autophagy,Brassica napus

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Characterization and mapping of a novel mutant sms1 (senescence and male sterility 1) in rice

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Journal of Genetics and Genomics 2010年第1期37卷 47-55页

作者： Wenyi Yan Shenghai Ye Qingsheng Jin Longjun Zeng Yu Peng Dawei Yan Weibing Yang Donglei Yang Zuhua He Yanjun Dong Xiaoming Zhang College of Life and Environment Sciences Shanghai Normal UniversityShanghai 200234China Institute of Crop and Nuclear Technology Utilization Zhejiang Academy of Agricultural SciencesHangzhou 310021China Institute of Plant Physiology & Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032China

Plant senescence plays diverse important roles in development and environmental ***,the molecular basis of plant senescence is remained largely unknown.A rice spontaneous mutant with the character of early senescence and male sterility （sms） was found in the breeding line *** order to identify the gene SMS1 and the underlying mechanism,we preliminarily analyzed physiological and biochemical phenotypes of the *** mutant contained lower chlorophyll content compared with the wild type control and was severe male sterile with lower pollen *** analysis showed that the mutant was controlled by a single recessive *** the map-based cloning approach,we fine-mapped SMS1 to a 67 kb region between the markers Z3-4 and Z1-1 on chromosome 8 using 1,074 F2 recessive plants derived from the cross between the mutant sms1 （japonica） × Zhenshan 97 （indica）,where no known gene involved in senescence or male sterility has been *** the SMS1 gene will be a novel gene that regulates the two developmental *** further cloning and functional analysis of the SMS1 gene is under way.

关键词： senescence male sterility rice molecular marker gene mapping

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Genetic Studies on Wheat Genic male sterility and Inheritance Mode of male sterility in Plant

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Agricultural Sciences in China 2000年第1期 24-30页

作者： Liu Binghua Institute of Crop Breeding and Cultivation Chinese Academy of Agricultural Science Crop Genetics and Breeding Key Lab of Nation Agriculture DepartmentBeijing 100081

Advance in the genetic study on male sterility in plant was reviewed. A dominant gene Ms2(Ta1) for male sterility, which hasa recombination percentage of 31. 16 with the centromere , is located on the short arm of chromosome 4D in Taigu wheat. Dwarfing male sterile wheat is a dwarf marked male sterile material, carrying the gene Ms2 for male sterility and dominant dwarf gene Rht10. The two genes, Ms2 and Rht10, are closely linked with a crossing-over value as small as 0. 18. The linkage character of genes Ms2 and Rht10 was transferred to CS ph1b mutant and octoploid triticale. A female-male sterility controlled by a set of genes and a male sterility carrying double recessive genes were discovered in wheat. The hypothesis of the origin of the male sterile form for gene interaction was proposed. An inheritance mode of fertility in sib-mating progenies of reces sive genic male sterile material in plant was established.

关键词： male sterility Genetic study Inheritance mode

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