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A reactive oxygen species burst causes haploid induction in maize

Molecular Plant 2022年第6期15卷 943-955页

作者： Chenglin Jiang Ju Sun Rui Li Shijuan Yan Wei Chen Liang Guo Guochen Qin Pengcheng Wang Cheng Luo Wenjie Huang Qinghua Zhang Alisdair R.Fernie David Jackson Xiang Li Jianbing Yan National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070China Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization Agro-biological Gene Research CenterGuangdong Academy of Agricultural SciencesGuangzhou 510640China Shanghai Center for Plant Stress Biology CAS Center for Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghai 200032China Department of Molecular Physiology Max-Planck-Institute of Molecular Plant PhysiologyAm Mühlenberg 114476 Potsdam-GolmGermany Cold Spring Harbor Laboratory Cold Spring HarborNY 11724USA State Key Laboratory of Plant Genomics and National Center for Plant Gene Research Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China Hubei Hongshan Laboratory Wuhan 430070China

haploid induction (HI) is an important tool in crop breeding. Phospholipase A1 (ZmPLA1)/NOT LIKE DAD (NLD)/MATRILINEAL (MTL) is a key gene controlling HI in maize;however, the underlying molecular mechanism remains unclear. In this study, to dissect why loss of ZmPLA1 function could mediate HI we performed a comprehensive multiple omics analysis of zmpla1 mutant anthers by integrating transcriptome, metabolome, quantitative proteome, and protein modification data. Functional classes of significantly enriched or differentially abundant molecular entities were found to be associated with the oxidative stress response, suggesting that a reactive oxygen species (ROS) burst plays a critical role in HI. In support of this, we further discovered that a simple chemical treatment of pollen with ROS reagents could lead to HI. Moreover, we identified ZmPOD65, which encodes a sperm-specific peroxidase, as a new gene controlling HI. Taken together, our study revealed a likely mechanism of HI, discovered a new gene controlling HI, and created a new method for HI in maize, indicating the importance of ROS balance in maintaining normal reproduction and providing a potential route to accelerate crop breeding.

关键词： haploid induction single-cell sequencing maize ROS peroxidase

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Construction of homozygous diploid potato through maternal haploid induction

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aBIOTECH 2022年第3期3卷 163-168页

作者： Jinzhe Zhang Jian Yin Jiayi Luo Die Tang Xijian Zhu Jie Wang Zhihong Liu Pei Wang Yu Zhong Chenxu Liu Canhui Li Shaojiang Chen Sanwen Huang Institute of Vegetables and Flowers Chinese Academy of Agricultural SciencesKey Laboratory of Biology and Genetic Improvement of Horticultural Crops of the Ministry of AgricultureSino-Dutch Joint Laboratory of Horticultural GenomicsBeijing100081China Shenzhen Branch Guangdong Laboratory of Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of Agriculture and Rural AffairsAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen518120China The AGISCAAS-YNNU Joint Academy of Potato Sciences Yunnan Normal UniversityKunming650500China National Maize Improvement Center of China Key Laboratory of Crop Heterosis and Utilization/Engineering Research Center for Maize BreedingMinistry of EducationCollege of Agronomy and BiotechnologyChina Agricultural UniversityBeijing100193China

Reinventing the tetraploid potato into a seed-propagated,diploid,hybrid potato would significantly accelerate potato *** this regard,the development of highly homozygous inbred lines is a prerequisite for breeding hybrid potatoes,but self-incompatibility and inbreeding depression present challenges for developing pure inbred *** resolve this impediment,we developed a doubled haploid(DH)technology,based on mutagenesis of the potato DOMAIN OF UNKNOWN FUNCTION 679 membrane protein(StDMP)***,we show that a deficiency in StDMP allows the generation of maternal haploids for generating diploid potato *** exercisable protocol,involving hybridization,fluorescent marker screening,molecular and flow cytometric identification,and doubling with colchicine generates nearly 100%homozygous diploid potato *** dmp-triggered haploid induction(HI)system greatly shortens the breeding process and offers a robust method for generating diploid potato inbred lines with high purity.

关键词： Doubled haploid technology haploid induction Diploid potato Inbred lines StDMP

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High temperature increases centromeremediated genome elimination frequency and enhances haploid induction in Arabidopsis

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Plant Communications 2023年第3期4卷 126-138页

作者： Ulkar Ahmadli Manikandan Kalidass Lucie Crhak Khaitova Joerg Fuchs Maria Cuacos Dmitri Demidov Sheng Zuo Jana Pecinkova Martin Mascher Mathieu Ingouff Stefan Heckmann Andreas Houben Karel Riha Inna Lermontova Leibniz Institute of Plant Genetics and Crop Plant Research(IPK)Gatersleben Corrensstrasse 306466 SeelandGermany Central European Institute of Technology(CEITEC)and National Centre for Biomolecular Research Faculty of ScienceMasaryk UniversityKamenice 562500 BrnoCzech Republic CIRAD DIADEIRDUniversity of Montpellier34393 MontpellierFrance

Double haploid production is the most effective way to create true-breeding lines in a single *** Arabidopsis,haploid induction via mutation of the centromere-specific histone H3(cenH3)has been shown when the mutant is outcrossed to the wild-type,and the wild-type genome remains in the haploid ***,factors that affect haploid induction are still poorly ***,we report that a mutant of the cenH3 assembly factor Kinetochore Null2(KNL2)can be used as a haploid inducer when pollinated by the *** discovered that short-term temperature stress of the knl2 mutant increased the efficiency of haploid induction *** also demonstrated that a point mutation in the CENPC-k motif of KNL2 is sufficient to generate haploid-inducing lines,suggesting that haploidinducing lines in crops can be identified in a naturally occurring or chemically induced mutant population,avoiding the generic modification(GM)approach at any ***,a cenh3-4 mutant functioned as a haploid inducer in response to short-term heat stress,even though it did not induce haploids under standard ***,we identified KNL2 as a new target gene for the generation of haploid-inducer lines and showed that exposure of centromeric protein mutants to high temperature strongly increases their haploid induction efficiency.

关键词： centromere kinetochore null 2 CENPC-k cenh3-4 temperature stress haploid induction

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A female in vivo haploid-induction system via mutagenesis of egg cell-specific peptidases

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Molecular Plant 2023年第2期16卷 471-480页

作者： Xuecheng Zhang Ce Shi Siling Li Bo Zhang Pan Luo Xiongbo Peng Peng Zhao Thomas Dresselhaus Meng-Xiang Sun State Key Laboratory of Hybrid Rice College of Life SciencesWuhan UniversityWuhan 430072China Hubei Hongshan Laboratory Wuhan 430070China Cell Biology and Plant Biochemistry University of RegensburgRegensburgGermany

Crop breeding schemes can be significantly accelerated by using(doubled)haploid *** vivo haploid induction has been applied in plant breeding for decades but is still not available for all crops and genotypes,and haploidization rates are generally very ***,methodological improvements to and new concepts for haploidization are ***,we report a novel system for the induction of haploid plants by mutating genes encoding egg cell-specific aspartic endopeptidases(ECSs).We show that after successful sperm–egg cell fusion,ECSs play a critical role to ensure male and female nucleus fusion after *** ecs1 ecs2 double mutant can induce haploids by both selfing and hybridization in Arabidopsis and ECS mutation is also capable of producing haploids in *** summary,our study develops a novel approach for maternal haploidization and provides new insights into the molecular basis of fertilization.

关键词： haploid induction egg cell maternal inducer semigamy fertilization

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Doubled haploid technology and synthetic apomixis:Recent advances and applications in future crop breeding

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Molecular Plant 2024年第7期17卷 1005-1018页

作者： Yanzhi Qu Alisdair R.Fernie Jie Liu Jianbing Yan National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070China Hubei Hongshan Laboratory Wuhan 430070China Department of Molecular Physiology Max-Planck-Institute of Molecular Plant Physiology14476 Potsdam-GolmGermany Center of Plant Systems Biology and Biotechnology 4000 PlovdivBulgaria azhouwan National Laboratory Sanya 572024China

Doubled haploid(DH)technology and synthetic apomixis approaches can considerably shorten breeding cycles and enhance breeding *** with traditional breeding methods,DH technology offers the advantage of rapidly generating inbred lines,while synthetic apomixis can effectively fix hybrid *** this review,we focus on(i)recent advances in identifying and characterizing genes responsible for haploid induction(Hl),(ii)the molecular mechanisms of Hl,(ili)spontaneous haploid genome doubling,and(iv)crop synthetic *** also discuss the challenges and potential solutions for future crop breeding programs utilizing DH technology and synthetic ***,we provide our perspectives about how to integrate DH and synthetic apomixis for precision breeding and de novo domestication.

关键词： maize doubled haploid technology haploid induction synthetic apomixis precision breeding

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AtRKD5 inhibits the parthenogenic potential mediated by AtBBM

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Journal of Integrative Plant Biology 2024年第7期66卷 1517-1531页

作者： Qiyan Liu Dongfen Han Denghu Cheng Jinfan Chen Shujuan Tian Jiafa Wang Man Liu Li Yuan State Key Laboratory of Crop Stress Resistance and High‐Efﬁciency Production College of HorticultureNorthwest A&F UniversityYangling 712100China

Parthenogenesis,the development of unfertilized egg cells into embryos,is a key component of ***(BABY BOOM),a crucial regulator of embryogenesis in Arabidopsis,possesses the capacity to shift nutritional growth toward reproductive ***,the mechanisms underlying AtBBM-induced parthenogenesis remain largely unexplored in dicot *** findings revealed that in order to uphold the order of sexual reproduction,the embryo-specific promoter activity of AtBBM as well as repressors that inhibit its expression in egg cells combine to limiting its ability to induce ***,AtRKD5,a RWPRK domain-containing(RKD)transcription factor,binds to the 3'end of AtBBM and is identified as one of the inhibitory factors for AtBBM expression in the egg *** the atrkd5 mutant,we successfully achieved enhanced ectopic expression of AtBBM in egg cells,resulting in the generation of haploid offspring via parthenogenesis at a rate of 0.28%.Furthermore,by introducing chimeric Arabidopsis and rice BBM genes into the egg cell,we achieved a significant 4.6-fold enhancement in haploid induction through the atdmp8/9 *** findings lay a strong foundation for further exploration of the BBM-mediated parthenogenesis mechanism and the improvement of haploid breeding efficiency mediated by the dmp8/9 mutant.

关键词： AtBBM atdmp AtRKD5 haploid induction parthenogenesis

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haploids can be induced in knockout mutants of OsPLA1,but not OsDMP3 or OsDMP6,in rice

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The Crop Journal 2024年第1期12卷 213-221页

作者： Zongkai Liu Yu Zhong Xiaolong Qi Tai An Shuwei Guo Dong Wang Yuwen Wang Bin Feng Zuofeng Zhu Shaojiang Chen Chenxu Liu National Maize Improvement Center Engineering Research Center of Ministry of EducationChina Agricultural UniversityBeijing 100193China National Center for Evaluation of Agricultural Wild Plants(Rice) Department of Plant Genetics and BreedingChina Agricultural UniversityBeijing 100193China

Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding ***1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,exhibiting a synergistic ***,it is unknown whether knock out of ZmDMP orthologs can stimulate HI in *** this study,a ZmPLA1 ortholog(OsPLA1)and two ZmDMP orthologs(OsDMP3 and OsDMP6)were identified in *** three genes encode plasma membrane-localized proteins and were highly expressed in mature *** of OsPLA1 in both Minghui 63 and Nipponbare resulted in reduced seed setting rate(SSR)and caused *** osdmp3,osdmp6 and the double mutant failed to trigger HI independently,nor increased the haploid induction rate(HIR)when combined with *** pollinations operations of QX654A with the ospla1 mutant significantly improve SSR,while reducing ***-seq profiling of mature ospla1 mutant anthers indicated that a large number of differentially expressed genes(DEGs)were enriched in redox homeostasis and lipid metabolic GO terms,plant hormone signal transduction,and MAPK signaling *** findings provide important insights towards construction of an efficient DH breeding technology and study of the molecular mechanism of HI in rice.

关键词： Doubled haploid breeding haploid induction Mutation anlaysis Rice

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The development and application of single cell sequencing technology in maize

The development and application of single cell sequencing te...

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2016年全国植物生物学大会

作者： Xiang Li Jianbing Yan National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University

Single cell sequencing technology emerged recently,enabling high-throughput analyses of the cell lineage trees of higher organisms. In plants,single-cell sequencing is still challenging because the cell wall hinders the isolation and lysis of the nuclear contents. We have developed a simple method to isolate and sequence the whole genome of each of the four microspores from a plant tetrad which was used to study the meiotic recombination mechanism of maize male gametes(Li et al,2015). Recently,we made further efforts to improve the method for isolating the three nuclei from one mature pollen grain of maize for whole-genome sequencing. This technique was used to study the haploid induction mechanism in maize. Production of maternal haploids via intra-specific genotypes as the haploid inducer is routine and highly efficient in maize. However,the underlying mechanism of haploid induction(HI) is unclear. We found that the aneuploidy caused by chromosome fragmentation occurring post meiosis in the gametophyte may affect embryogenesis and be the underlying cause of HI.

关键词： single cell sequencing recombination haploid induction chromosome fragmentation

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