QTL mapping of grain appearance quality traits and grain weight using a recombinant inbred population in rice(Oryza sativa L.)

supported by a grant from the National High-Tech R＆D Program of China （2014AA10A603, 2014AA10A604）;a grant from the Youth Foundation in Sichuan, China （2011JTD0022）;the special fund for China Agricultural Research System （CARS-01-08）;the Provincial Specialized Funds for Innovation Ability Promotion in Sichuan, China （2013GXJS005）

Comparison of Morphological and Genetic Differentiations in Filial Generation of Cross Between Indica and Japonica Rice

A recombinant inbred line （RIL） population of F8 and F9 generations derived from a cross between a typical indica rice （Qishanzhan） and a typical japonica rice （Akihikari） was used to study the difference between morphological differentiation based on phenotype characters and genetic differentiation using indica and japonica specific SSR markers, and to evaluate the relationship between vascular bundle characters and morphological and genetic differentiations. The results showed that the frequency distributions of morphological and genetic differentiations were all inclined to japonica type in the filial generation. The population was more inclined to japonica type based on genetic differentiation than on morphological differentiation. The consistent degrees of classification based on the Cheng’s index, the ratio of large vascular bundle number to small vascular bundle number in panicle neck （RLSVB） and the ratio of large vascular bundle number in the second internode from the top to that in the panicle neck （RLVB） were all about 50% compared with the genetic differentiation, and the consistent degree of the total scores of the Cheng’s index combined with the vascular bundle number ratios was significantly increased to about 80% compared with the genetic differentiation. Therefore, the vascular bundle characters could be used as a helpful supplement for subspecies classification.

Effect of Environment and Genetic Recombination on Subspecies and Economic Trait Differentiation in the F_2 and F_3 Generations from indicajaponica Hybridization

indica and japonica are the two most important subspecies of Asian cultivated rice. Identifying mechanisms responsible for population differentiation in these subspecies is important for indica-japonica hybridization breeding. In this study, subspecies and economic trait differentiation patterns were analyzed using morphological and molecular （InDel and Intron Length Polymorphism） data in F2 and F3 populations derived from indica-japonica hybridization. Populations were grown in Liaoning and Guangdong provinces, China, with F3 populations generated from F2 populations using bulk harvesting （BM） and single-seed descent methods （SSD）. Segregation distortion was detected in F3-BM populations, but not in F3- SSD or in F2 populations. Superior performance was observed with respect to economic traits in Liaoning compared with that in Guangdong and 1 000-grain weight （KW）, seed setting rate （SSR） and grain yield per plant （GYP） were significantly correlated with indica and japonica subspecies types. Analysis of molecular and morphological data demonstrated that the environment is the main factor giving rise to population differentiation in indica-japonica hybridization. In addition, we also found that KW, SSR and GYP are related to subspecies characteristics and kinship, which is possibly a significant factor resulting in economic trait differentiation and determining environmental adaptability. Our study has provided new insights into the process of population differentiation in these subspecies to inform indica-japonica hybridization breeding.

TALEN-Mediated Homologous Recombination Produces Site-Directed DNA Base Change and Herbicide-Resistant Rice

Over the last decades,much endeavor has been made to advance genome editing technology due to its promising role in both basic and synthetic biology.The breakthrough has been made in recent years with the advent of sequence-specific endonucleases,especially zinc finger nucleases（ZFNs）,transcription activator-like effector nucleases（TALENs） and clustered regularly interspaced short palindromic repeats（CRISPRs） guided nucleases（e.g.,Cas9）.In higher eukaryotic organisms,site-directed mutagenesis usually can be achieved through non-homologous end-joining（NHEJ） repair to the DNA double-strand breaks（DSBs） caused by the exogenously applied nucleases.However,site-specific gene replacement or genuine genome editing through homologous recombination（HR） repair to DSBs remains a challenge.As a proof of concept gene replacement through TALEN-based HR in rice（Oryza sativa）,we successfully produced double point mutations in rice acetolactate synthase gene（OsALS） and generated herbicide resistant rice lines by using TALENs and donor DNA carrying the desired mutations.After ballistic delivery into rice calli of TALEN construct and donor DNA,nine HR events with different genotypes of OsALS were obtained in T_0 generation at the efficiency of 1.4%—6.3%from three experiments.The HRmediated gene edits were heritable to the progeny of T_1 generation.The edited T_1 plants were as morphologically normal as the control plants while displayed strong herbicide resistance.The results demonstrate the feasibility of TALEN-mediated genome editing in rice and provide useful information for further genome editing by other nuclease-based genome editing platforms.