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| 1 | Target chromosome-segment substitution: A way to breeding by design in rice显示文摘Progress in plant breeding depends on the development of genetic resources,genetic knowledge,and breeding techniques.The core of plant breeding is the use of naturally occurring variation.At the beginning of the post-genomic era,a new concept of'breeding by design'was proposed,which aims to control all allelic variation for all genes of agronomic importance.In the past two decades,we have applied a three-step strategy for research on rice breeding by design.In the first step,we constructed a singlesegment substitution line(SSSL)library using Huajingxian 74(HJX74),an elite xian(indica)rice cultivar,as the recipient in which to assemble genes from the rice AA genome.In the second step,we identified a series of desirable genes in the SSSL library.In the third step,we designed new rice lines,and achieved the breeding goals by pyramiding target genes in the HJX74-SSSL library.This review introduces the background,concept,and strategy of breeding by design,as well as our achievements in rice breeding by design using the HJX74-SSSL platform.Our practice shows that target chromosome-segment substitution is a way to breeding by design. | Guiquan Zhang | 2021 | The Crop Journal2021，9，3： | 5 |
| 2 | Establishment of an efficient seed fluorescence reporter-assisted CRISPR/Cas9 gene editing in maize显示文摘Genome editing by clustered regularly interspaced short palindromic sequences(CRISPR)/CRISPRassociated protein 9(Cas9)has revolutionized functional gene analysis and genetic improvement.While reporter-assisted CRISPR/Cas systems can greatly facilitate the selection of genome-edited plants produced via stable transformation,this approach has not been well established in seed crops.Here,we established the seed fluorescence reporter(SFR)-assisted CRISPR/Cas9 systems in maize(Zea mays L.),using the red fluorescent Ds RED protein expressed in the endosperm(En-SFR/Cas9),embryos(Em-SFR/Cas9),or both tissues(Em/En-SFR/Cas9).All three SFRs showed distinct fluorescent patterns in the seed endosperm and embryo that allowed the selection of seeds carrying the transgene of having segregated the transgene out.We describe several case studies of the implementation of En-SFR/Cas9,Em-SFR/Cas9,and Em/En-SFR/Cas9 to identify plants not harboring the genomeediting cassette but carrying the desired mutations at target genes in single genes or in small-scale mutant libraries,and report on the successful generation of single-target mutants and/or mutant libraries with En-SFR/Cas9,Em-SFR/Cas9,and Em/En-SFR/Cas9.SFR-assisted genome editing may have particular value for application scenarios with a low transformation frequency and may be extended to other important monocot seed crops. | Yuanyuan Yan Jinjie Zhu Xiantao Qi Beijiu Cheng Changlin Liu Chuanxiao Xie | 2021 | Journal of Integrative Plant Biology2021，63，9： | 3 |
| 3 | 西南稻区重穗型杂交稻发展现状及分子设计育种策略显示文摘水稻是我国重要的粮食作物,全国有60%以上人口以大米为主食。西南稻区是我国六大稻区之一,对保障我国粮食安全具有重要的战略意义。针对西南稻区“寡日照、高湿度、小温差”的生态条件,周开达院士等经历了艰苦的探索和反复实践,提出发展“重穗型”杂交稻是西南稻区实现水稻超高产的重要途径。本文总结了西南稻区重穗型杂交稻发展现状,列举了可用于重穗型杂交稻分子设计育种的基因资源,提出了重穗型杂交稻的分子设计育种策略,以期为培育新的高产优质“重穗协调型”品种提供参考,致力于打造更高水平的“天府粮仓”。 | 袁华 陈薇兰 王淏 王玉平 马炳田 涂斌 钦鹏 李仕贵 | 2022 | 中国稻米2022，28，5： | 2 |
| 4 | 基于原生质体的谷子CRISPR/Cas9基因编辑系统优化显示文摘为了优化和筛选谷子中高效的CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats/CRISRP-associated nuclease 9)基因编辑系统,针对谷子八氢番茄红素脱氢酶基因(SiPDS)设计6种gRNA(gRNA1—gRNA2针对外显子1,gRNA3—gRNA6针对外显子12),构建多种CRISPR/Cas9基因编辑系统,通过聚乙二醇(PEG)介导的方法转入谷子原生质体中,然后利用开发的大规模平行测序技术快速检测它们对SiPDS的突变效率。结果表明,采用7 d谷子黄化苗幼茎建立的原生质体转化系统的转化效率较高,为50.44%~57.36%。将Super启动子(SP)、谷子内源的泛素启动子(Ubi)分别驱动Cas9基因的基因编辑系统转入谷子原生质体中,发现两者对SiPDS基因的突变效率分别为0.5%、5.5%。随后采用Ubi启动子驱动Cas9基因,比较单一gRNA、双gRNA和tRNA结构的基因编辑系统对SiPDS基因的突变效率,发现双gRNA基因编辑系统Ubi-dgRNAE1和Ubi-dgRNAE12的突变效率分别较单一gRNA基因编辑系统提高了1.66倍和1.11倍;tRNA基因编辑系统Ubi-tRNA的突变效率较单一gRNA基因编辑系统提高了5.87倍,为51.24%,且Ubi-tRNA可同时针对多个位点进行编辑,其多位点突变频率为3.23%。比较gRNA3、gRNA4、gRNA5体外转录产物分别与Cas9蛋白混合制成的核糖核蛋白(RNP)复合物的体外切割活性,发现RNP-gRNA5复合物体外切割活性最高,其对SiPDS基因的突变效率是2.0%,并且引起的突变类型主要为小于3 bp的缺失。 | 刘光宇 徐晓静 夏科科 孙海汐 陶月如 崔震 顾颖 | 2022 | 河南农业科学2022，51，1： | 0 |
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