OsPPR9 encodes a DYW-type PPR protein that affects editing efficiency of multiple RNA editing sites and is essential for chloroplast development

Photosynthesis occurs mainly in chloroplasts,whose development is regulated by proteins encoded by nuclear genes.Among them,pentapeptide repeat(PPR)proteins participate in organelle RNA editing.Although there are more than 450 members of the PPR protein family in rice,only a few affect RNA editing in rice chloroplasts.Gene editing technology has created new rice germplasm and mutants,which could be used for rice breeding and gene function study.This study evaluated the functions of OsPPR9 in chloroplast RNA editing in rice.The osppr9 mutants were obtained by CRISPR/Cas9,which showed yellowing leaves and a lethal phenotype,with suppressed expression of genes associated with chloroplast development and accumulation of photosynthetic-related proteins.In addition,loss of OsPPR9 protein function reduces the editing efficiency of rps8-C182,rpoC2-C4106,rps14-C80,and ndhB-C611 RNA editing sites,which affects chloroplast growth and development in rice.Our data showed that OsPPR9 is highly expressed in rice leaves and encodes a DYW-PPR protein localized in chloroplasts.Besides,the OsPPR9 protein was shown to interact with OsMORF2 and OsMORF9.Together,our findings provide insights into the role of the PPR protein in regulating chloroplast development in rice.

水稻苗期根部性状的遗传分析和最长根长QTL qLRL4的精细定位

为了解析水培苗期根系相关性状的遗传调控,以籼稻9311和粳稻日本晴(Nipponbare, NPB)为亲本的148个株系构成的重组自交系群体为材料,对水稻幼苗根系相关性状开展QTL分析。在2次重复中共检测到26个控制最长根长、总根系长、根表面积、根体积和根直径的QTL,分布在水稻第1、2、4、7、9、10、11号共7条染色体上,发现了水稻第2、4、7和10号染色体上的4个QTL簇,包括第4号染色体上控制最长根长的QTL qLRL4。为了精细定位该QTL,我们构建了以9311为背景、插入缺失标记IND4-1和IND4-4间来自NPB的近等系NIL-qLRL4。利用NIL-qLRL4和9311构建的F2群体,最终将qLRL4精细定位在标记IND4-1和IND4-3之间约68.23 kb的区间内并预测了候选基因。此根长QTL的精细定位将有助于水稻根长遗传机理的研究,为探究水稻根系形态建成的分子机制奠定了基础。

Use of Major Quantitative Trait Loci to Improve Grain Yield of Rice

Further improvement of rice productivity remains a challenge. Breeding is perceived as an important option to increase rice yield. However, the genetic progress of grain yield in most rice breeding programs was slow in the last decades. Although great progress in rice genomics and molecular biology has been achieved, the effect of such technological innovations on rice breeding is far small. Marker-assisted selection （MAS） for a few target quantitative trait loci （QTLs） has significant effects in improving qualitative traits, such as disease resistance. The success of MAS has therefore motivated breeders to identify and use major QTLs for yield and yield component traits. In this review, we summarized the recent methods in QTL identification, including novel statistical methods for linkage and association mapping, special population types, and whole-genome sequencing. We reviewed the successful application of marker-assisted gene introgression and gene pyramiding to improve grain yield and discussed the design of efficient MAS schemes to further increase the success rate of breeding programs. The use of well-characterized major QTLs through introgression and gene pyramiding is proven effective in improving grain yield, particularly yield under abiotic stress. Major QTLs that are stable across genetic background and growing environments are often found in less adapted germplasms, such as landraces and wild relatives. Advanced backcross QTL analysis and introgression lines, which integrate QTL discovery and utilization, are important methods for exploiting major QTLs contained in such germplasms. Next-generation sequencing substantially increases mapping resolution and accelerates the identification of casual genes underlying major QTLs. Practical guidelines derived from theoretical and empirical studies are given to guide the design of efficient marker-assisted gene introgression and pyramiding schemes.

Molecular Improvement of Grain Weight and Yield in Rice by Using GW6 Gene

Molecular design breeding is one of straightforward approaches to break yield barriers in rice. In this study, GW6 gene for grain length and width from Baodali was transferred into an indica recurrent parent 9311 and a japonica variety Zhonghua 11 （ZH11） using marker-assisted backcross （MAB）. One and three introgression lines were selected for phenotypic analysis from 9311 and ZH11 genetic backgrounds, respectively. SSL-1, an improved 9311 near isogenic line with GW6 performed 11%, 19% and 6.7%higher of grain length, 1000-grain weight and single plant yield, respectively, as compared with 9311. All the three improved ZH11-GW6 lines, R1, R2 and R3, had more than 30% increase in grain weight and about 7%higher in grain yield. Seed plumpness of R1, R2 and R3 was improved synchronously because the three ZH11-GW6 lines contained GIF1 （Grain Incomplete Filling 1）, a dominant grain filling gene. Thus, GW6 has high potential in increasing the yield of inbred lines through MAB, making it an important genetic resource in super hybrid rice breeding. This study provides insights in the utilization of GW6 for large grain and high yield rice breeding via molecular design breeding.

A Simple Method for Preparation of Rice Genomic DNA

The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection （MAS） programs. A simple method for preparation of rice genomic DNA was developed. A small amount （1~50 mg） of leaf tissue of rice seedling, 500 pL of extraction buffer, and one steel bead were put into a 2-mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of supernatant was directly applied to PCR amplification. Otherwise, the supematant was precipitated with two times volume of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 min. It is especially suitable for genotyping of large number of samples.

Genetic Analysis and QTL Mapping of Mature Seed Culturability in Indica Rice

Genetic segregation analysis for mature seed culturability was conducted using recombinant inbred lines derived from a cross of indica rice, Yangdao 6 and Pei＇ai 64s. Three indices of seed culturability, the frequency of callus induction, the frequency of brown callus and the increase of callus weight were investigated. A combined genetic map constructed with simple sequence repeat （SSR）, sequence tag site （STS）, cleaved amplified polymorphic sequences （CAPS） and single nucleotide polymorphism （SNP） markers covered a total distance of 1 732.5 cM, averaging approximately 12 cM between two neighboring loci. Three QTLs on chromosomes 7, 7 and 10 were detected for the frequency of callus induction; three QTLs on chromosomes 6, 7 and 9 were detected for the frequency of brown callus; and two QTLs on chromosomes 5 and 7 were detected for the increase of callus weight. Common QTLs mapped at the interval flanking RM5481 and RM6835 on chromosome 7 were identified to be involved in the frequency of callus induction and the frequency of brown callus, explaining 7.29% and 12.52% of phenotypic variation, respectively. A total of 14 epistatic effects were detected for the three indices of mature seed culturability. ~

Characterization and Fine Mapping of Non-panicle Mutant (nop) in Rice

A mutant of panicle differentiation in rice called non-panicle （nop） was discovered in the progeny of a cross between 93-11 and Nipponbare. The mutant exhibits normal plant morphology but has apparently few tillers. The most striking change in nop is that its panicle differentiation is blocked, with masses of fluffy bract nodes generate from the positions where rachis branches normally develop in wild-type plants. Genetic analysis suggests that nop is controlled by a single recessive gene, which is temporarily named Nop（t）. Based on its mutant phenotype, Nop（t） represents a key gene controlling the initiation of inflorescence differentiation, By using simple sequence repeat markers and sequence tagged site markers, Nop（t） gene was fine mapped in a 102-kb interval on the long arm of chromosome 6. These results will facilitate the positional cloning and functional studies of the gene.