Genetic diversity center of cultivated soybean(Glycine max) in China——New insight and evidence for the diversity center of Chinese cultivated soybean

Information on the center of genetic diversity of soybean（Glycine max） will be helpful not only for designing efficient strategies for breeding programs, but also for understanding the domestication and origin of this species. Here, we describe an analysis of genetic diversity based on simple-sequence repeat（SSR） variations within a core collection of 2 111 accessions of Chinese soybean landraces. Prior to the diversity assessment, the geographic origin of each accession was mapped. The map was then divided into grids each 2.5° in latitude and 5° in longitude. We found two regions that had higher number of alleles（NA） and greater polymorphic information content（PIC） values than the others. These regions are adjacently located within grid position of 30°–35°N×105°–110°E, which includes the valley of the middle and lower reaches of the Wei River, and the valley of the upper reaches of the Hanjiang River. It was also observed that in many regions, genetic diversity decreased with the increase in distance from the center. Another region, in northern Hebei Province（115°–120°E×40°–42.5°N）, was observed having higher diversity than any surrounding regions, indicating that this is a sub-center of soybean diversity. Based on the presented results, the domestication and origin of soybean are also discussed.

Analysis of Genetic Similarity for Improved Japonica Rice Varieties from Different Provinces and Cities in China

To provide a genetic basis for japonica rice breeding, the genetic similarity and cluster of 139 accessions of improved japonica rice varieties from 12 provinces and cities of China were analyzed using 34 SSR markers. Totally 198 alleles were detected among these improved japonica rice varieties with the average number of alleles per pair of primers was 5.3235. RM320, RM531, RM1, RM286, and RM336 showed more alleles, which were 15, 12, 11, 9, and 9, respectively. RM320, RM336, RM286 and RM531 showed higher genetic diversity indexes; which were 2.3324, 2.0292, 1.8996, and 1.7820, respectively. The range of genetic similar index among improved japonica rice varieties from different provinces was from 0.321 to 0.914, with the average of 0.686. There was a high genetic similarity among improved japonica rice varieties from Heilongjiang, Jilin, Liaoning, Ningxia, and Yunnan, which were located in similar latitude or similar ecological environment, while there was a low genetic similarity between improved japonica rice varieties from Guizhou and Jiangsu, and other provinces which were located in more different latitudes and ecological environments. The markers of RM320, RM531, RM1, RM286, and RM336 fit to be used in analysis of genetic diversity for improved japonica rice variety. The genetic similarity among improved japonica rice varieties from different provinces was closely associated with genetic basis of parents, and was also correlated with latitude and ecological environment where the varieties were bred.

Integrated genomic and transcriptomic analysis reveals genes associated with plant height of foxtail millet

Foxtail millet(Setaria italica)is an important C4 model crop;however,due to its high-density planting and high stature,lodging at the filling stage resulted in a serious reduction in yield and quality.Therefore,it is imperative to identify and deploy the genes controlling foxtail millet plant height.In this study,we used a semi-dwarf line 263A and an elite high-stalk breeding variety,Chuang 29 to construct an F2 population to identify dwarf genes.We performed transcriptome analysis(RNA-seq)using internode tissues sampled at three jointing stages of 263A and Chuang 29,as well as bulk segregant analysis(BSA)on their F2 population.A total of 8918 differentially expressed genes(DEGs)were obtained from RNA-seq analysis,and GO analysis showed that DEGs were enriched in functions such as‘‘gibberellin metabolic process”and‘‘oxidoreductase activity”,which have previously been shown to be associated with plant height.A total 593 mutated genes were screened by BSA-seq method.One hundred and seventy-six out of the 593 mutated genes showed differential expression levels between the two parental lines,and seven genes not only showed differential expression in two or three internode tissues but also showed high genomic variation in coding regions,which indicated they play a crucial role in plant height determination.Among them,we found a gibberellin biosynthesis related GA20 oxidase gene(Seita.5G404900),which had a single-base at the third exon,leading to the frameshift mutation at 263A.Cleaved amplified polymorphic sequence assay and association analysis proved the single-base in Seita.5G404900 co-segregated with dwarf phenotype in two independent F2 populations planted in entirely different environments.Taken together,the candidate genes identified in this study will help to elucidate the genetic basis of foxtail millet plant height,and the molecular marker will be useful for marker-assisted dwarf breeding.

The prevalence of deleterious mutations during the domestication and improvement of soybean

Soybean(Glycine max L.)is a protein and oil crop grown worldwide.Its fitness may be reduced by deleterious mutations,whose identification and purging is desirable for crop breeding.In the published whole-genome re-sequenced data of 2214 soybean accessions,including 221 wild soybean,1132 landrace cultivars and 861 improved soybean lines,we identified 115,275 deleterious single-nucleotide polymorphisms(SNPs).Numbers of deleterious alleles increased from wild soybeans to landraces and decreased from landraces to modern improved lines.Genes in selective-sweep regions showed fewer deleterious mutations than the remaining genes.Deleterious mutations explained 4.3%-48%more phenotypic variation than randomly selected SNPs for resistance to soybean cyst nematode race 2(SCN2),soybean cyst nematode race 3(SCN3)and soybean mosaic virus race 3(SMV3).These findings illustrate how mutation load has shifted during soybean domestication,expansion and improvement and provide candidate sites for breeding out deleterious mutations in soybean by genome editing and/or conventional breeding focused on the selection of progeny with fewer deleterious alleles.

Development of Soybean EST-SSR Markers and Their Use to Assess Genetic Diversity in the Subgenus Soja

Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA library. Among the 286 markers designed for the 4 accessions of Glycine max and 6 of its wild progenitor （G. soja） within the subgenus Soja, 209 markers amplified DNA fragments, taking 73.1% and 37 markers appeared to be polymorphic, which was 12.9% of the total. The 37 loci detected a total of 142 alleles, while the PIC values varied from 0.194 to 0.794. Both the number of alleles per locus and PIC value were significantly related to the SSR motif. Six EST-SSR loci may be fixed for different alleles between G. max and G. soja since they were particularly polymorphic among the 6 G. soja accessions. A neighbor-joining tree placed the G. max accessions together as a group within the G. soja, though the average genetic distance among G. soja accessions was much higher. These new EST-SSRs markers will be useful for genetic diversity analysis, genetic mapping construction and gene discovery in Soja subgenus.

Analysis of SSRs Uncovers Hierarchical Structure and Genetic Diversity in Chinese Soybean Landraces

For clarifying the hierarchical patterns of population structure of soybean landraces in China, the seven clusters previously identified using Bayesian clustering of 1 504 soybean landraces based on SSR markers genotyping data were further analyzed. Using the largest value of AK, these landraces could be split into 20 sub-clusters, which was supported by highly significant pairwise Fst-values and generally in accordance with the geographic origin and sowing types. The autumn-sowing types ended up in one distinct sub-cluster from the otherwise summer-sowing type, where the autumn- sowing types are most likely derived from. The division into 20 sub-clusters explained 7.3% of the genetic variation, next to 9.7% present among the seven clusters, 81.1% residing among landraces within sub-clusters, and 1.9% within the landraces. The distribution pattern of genetic diversity among the sub-clusters of each cluster was uneven, with two HSuM sub-clusters （Central China） and some South China sub-clusters showing significantly higher level of genetic diversity.

Mapping and validation of a dominant salt tolerance gene in the cultivated soybean(Glycine max) variety Tiefeng 8

Salt is an abiotic stress factor that strongly affects soybean growth and production. A single dominant gene has been shown to confer salt tolerance in the soybean cultivar Tiefeng 8.The objective of the present study was to genetically map the salt-tolerance gene in an F2:3population and a recombinant inbred line(RIL) population derived from a cross between two cultivated soybeans, Tiefeng 8(tolerant) and 85-140(sensitive). The F2:3families and RILs were treated with 200 mmol L-1Na Cl to evaluate salt tolerance. The F2:3population showed 1(42 tolerant): 2(132 segregating): 1(65 sensitive) segregation, indicating a single dominant gene for salt tolerance in Tiefeng 8. A sequence-characterized amplified region(SCAR) marker from a previously identified random amplified polymorphic DNA(RAPD)marker and four insertion/deletion polymorphism(In Del) markers were developed within the mapping region. Using these markers along with SSR markers, the salt-tolerance gene was mapped within 209 kb flanked by SCAR marker QS08064 and SSR marker Barcsoyssr_3_1301 on chromosome 3. Three markers that cosegregated with the salt tolerance gene and SCAR marker QS08064 were used to genotype 35 tolerant and 23 sensitive soybean accessions. These markers showed selection efficiencies of 76.2% to94.2%. The results indicate that these markers will be useful for marker-assisted breeding and facilitating map-based cloning of the salt tolerance gene in soybean.

HrcQ is necessary for Xanthomonas oryzae pv. oryzae HR-induction in non-host tobacco and pathogenicity in host rice

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice(Oryza sativa L.) worldwide. The type III secretion system(T3SS) of Xoo, encoded by the hrp(hypersensitive response and pathogenicity) genes, plays critical roles in conferring pathogenicity in host rice and triggering a hypersensitive response(HR) in non-host plants. To investigate the major genes conferring the pathogenicity and avirulence of Xoo, we previously constructed a random Tn5-insertion mutant library of Xoo strain PXO99A. We report here the isolation and characterization of a Tn5-insertion mutant PXM69. Tn5-insertion mutants were screened on indica rice JG30, which is highly susceptible to PXO99A, by leaf-cutting inoculation.Four mutants with reduced virulence were obtained after two rounds of screening. Among them, the mutant PXM69 had completely lost virulence to the rice host and ability to elicit HR in non-host tobacco. Southern blotting analysis showed a single copy of a Tn5-insertion in the genome of PXM69. PCR walking and sequencing analysis revealed that the Tn5 transposon was inserted at nucleotide position 70,192–70,201 in the genome of PXO99A, disrupting the type III hrc(hrp-conserved) gene hrcQ, the first gene in the D operon of the hrp cluster in Xoo. To confirm the relationship between the Tn5-insertion and the avirulence phenotype of PXM69, we used the marker exchange mutagenesis to create a PXO99Amutant, ΔhrcQ::KAN, in which the hrcQ was disrupted by a kanamycin-encoding gene cassette at the same site as that of the Tn5-insertion. ΔhrcQ::KAN showed the same phenotype as mutant PXM69. Reintroduction of the wild-type hrcQ gene partially complemented the pathogenic function of PXM69. RT-PCR and cellulase secretion assays showed that the Tn5-disruption of hrcQ did not affect transcription of downstream genes in the D operon and function of the type II secretion system. Our results provide new insights into the pathogenic functions of clustered hrp genes in Xoo.

Co-treatment with surfactant and sonication signifi cantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

Soybean is a widely planted genetically modified crop around the world. However, it is still one of the most recalcitrant crops for genetic transformation due to the difficulty of regeneration via organogenesis and some factors that affect the transformation efficiency. The percentages of resistant bud formation and transient expression efficiency are important indexes reflecting the regeneration and transformation efficiency of soybean. In this study, the percentages of resistant bud formation and transient expression of 13-glucuronidase （GUS） were compared after treatment with sonication or surfactant and co-treatment with both. The results showed that treatment with either sonication or surfactant increased the percentage of resistant bud formation and transient expression efficiency. The highest percentages were acquired and significantly improved when cotyledon node explants were co-treated with sonication for 2 s and surfactant at 0.02% （v：v） using two different soybean genotypes, Jack and Zhonghuang 10. The improved transformation efficiency of this combination was also evaluated by development of herbicide-tolerant soybeans with transformation efficiency at 2.5-5.7% for different geno- types, which was significantly higher than traditional cotyledonary node method in this study. These results suggested that co-treatment with surfactant and sonication significantly improved the percentages of resistance bud formation, transient expression efficiency and stable transformation efficiency in soybean.

Pseudomonas sp.TK35-L enhances tobacco root development and growth by inducing HRGPnt3 expression in plant lateral root formation

Rhizosphere colonization is a key requirement for the application of plant growth-promoting rhizobacteria(PGPR)as a bioferilizer.Signaling molecules are often exchanged between PGPR and plants,and genes in plants may respond to the action of PGPR.Here,the luciferase luxAB gene was electrotransformed into Pseudomonas sp.strain TK35,a PGPR with an afinity for tobacco,and the labelled TK35(TK35-L)was used to monitor colonization dynamics in the tobacco rhizosphere and evaluate the effects of colonization on tobacco growth and root development.The transcript levels of the hydroxyproline rich glycoprotein HRGPnt3 gene,a lateral root induction indicator,in tobacco roots were examined by qPCR.The results showed that TK35-L could survive for long periods in the tobacco rhizosphere and colonize new spaces in the tobacco rhizosphere following tobacco root extension,exhibiting significant increases in root development,seedling growth and potassium accumulation in tobacco plants.The upregulation of HRGPnt3 transcription in the inoculated tobacco suggested that TK35-L can promote tobacco root development by upregulating the transcript levels of the HRGPnt3 gene,which promotes tobacco seedling growth.These findings lay a foundation for future studies on the molecular mechanism underlying the plant growth-promoting activities of PGPR.Futhermore,this work provided an ideal potential strain for biofertilizer production.

Comparative Analysis of Gene Expression Profiling Between Resistant and Susceptible Varieties Infected With Soybean Cyst Nematode Race 4 in Glycine max

Soybean cyst nematode （SCN） is one of the most devastating pathogen for soybean. Therefore, identiifcation of resistant germplasm resources and resistant genes is needed to improve SCN resistance for soybean. Soybean varieties Huipizhiheidou and Wuzhaiheidou were distributed in China and exhibited broad spectrums of resistance to various SCN races. In this study, these two resistant varieties, combined with standard susceptible varieties （Lee and Essex）, were utilized to identify the differentially expressed transcripts after infection with SCN race 4 between resistant and susceptible reactions by using the Affymetrix Soybean Genome GeneChip. Comparative analyses indicated that 21 common genes changed signiifcantly in the resistant group, of which 16 increased and 5 decreased. However, 12 common genes changed signiifcantly in the susceptible group, of which 9 increased and 3 decreased. Additionally, 27 genes were found in common between resistant and susceptible reactions. The 21 signiifcantly changed genes in resistant reaction were associated with disease and defense, cell structure, transcription, metabolism, and signal transduction. The fold induction of 4 from the 21 genes was conifrmed by quantitative RT-PCR （qRT-PCR） analysis. Moreover, the gene ontology （GO） enrichment analyses demonstrated the serine family amino acid metabolic process and arginine metabolic process may play important roles in SCN resistance. This study provided a new insight on the genetic basis of soybean resistance to SCN race 4, and the identiifed resistant or resistant-related genes are potentially useful for SCN-resistance breeding in soybean.

Construction of Two Suppression Subtractive Hybridization Libraries and Identification of Salt-Induced Genes in Soybean

Soybean is planted worldwide and its productivity is significantly hampered by salinity. Development of salt tolerant cultivars is necessary for promoting soybean production. Despite wealth of information generated on salt tolerance mechanism, its basics still remain elusive. A continued effort is needed to understand the salt tolerance mechanism in soybean using suitable molecular tools. To better understand the molecular basis of the responses of soybean to salt stress and to get an enrichment of critical salt stress responsive genes in soybean, suppression subtractive hybridization libraries （SSH） are constructed for the root tissue of two cultivated soybean genotypes, one was tolerant and the other was sensitive to salt stress. To compare the responses of plants in salt treatment and non-treatment, SSH1 was constructed for the salt-tolerant cultivar Wenfeng 7 and SSH2 was constructed for the salt-sensitive cultivar Union. From the two SSH cDNA libraries, a total of 379 high quality ESTs were obtained. These ESTs were then annotated by performing sequence similarity searches against the NCBI nr （National Center for Biotechnology Information protein non-redundant） database using the BLASTX program. Sixty-three genes from SSH1 and 49 genes from SSH2 could be assigned putative function. On the other hand, 25 ESTs of SSH1 which may be not the salt tolerance-related genes were removed by comparing and analyzing the ESTs from the two S SH libraries, which increased the proportion of the genes related to salt tolerance in S SH 1. These results suggested that the novel way could realize low background of SSH and high level enrichment of target cDNAs to some extent.

Establishment and application of an accurate identification method for fragrant soybeans

In order to screen the aroma characteristics of soybean,a new method was established which can quickly quantify the content of 2-acetyl-1pyrroline(2-AP),an important compound related to soybean aroma,using gas chromatography-mass spectrometry(GC-MS).Based on peak profile,total peak area and retention time as test indexes,an accurate identification method for fragrant soybeans was established.The optimum parameters of the protocol consisted of column temperature70℃,sample injector temperature 180℃,optimum extraction alcohol content 1 m L,Na Cl content 0.1 g,ultrasonication time10 min,and extraction time 1 h,which were established by using the orthogonal test of single factors and three factors with four levels(L_9(3)^4).2-AP content of leaves had significant correlations with seeds,which were easier to measure.The protocol was simple and easy to carry out,consumed only small amounts of reagents,and provided accurate and reliable results with good reproducibility.A total of 101 soybean genotypes from different geographical sources were analyzed using this protocol.The results showed that the average content of 2-AP was 0.29 mg L^(–1),ranging from 0.094 to 1.816 mg L^(–1),and the genetic diversity index was 0.54.Among all genotypes-tested,they were classified into three grades,including seven elite genotypes identified as"grade one fragrant soybeans",which were Zhonglong 608,Heinong 88,Ha13-2958,Hongmiandou,Heinong 82,Huangmaodou,and Jiyu 21.These results provide both an identification technique and several elite aroma genotypes for gene discovery and good quality breeding in soybean.

Natural variation of GmFNSII-2 contributes to drought resistance by modulating enzyme activity in soybean

As an essential crop that provides vegetable oil and protein,soybean(Glycine max(L.)Merr.)is widely planted all over the world.However,the scarcity of water resources worldwide has seriously impacted on the quality and yield of soybean.To address this,exploring excellent genes for improving drought resistance in soybean is crucial.In this study,we identified natural variations of GmFNSII-2(flavone synthase II)significantly affect the drought resistance of soybeans.Through sequence analysis of GmFNSII-2 in 632 cultivated and 44 wild soybeans nine haplotypes were identified.The full-length allele GmFNSII-2^(C),but not the truncated allele GmFNSII-2^(A) possessing a nonsense nucleotide variation,increased enzyme activity.Further research found that GmDREB3,known to increase soybean drought resistance,bound to the promoter region of GmFNSII-2^(C).GmDREB3 positively regulated the expression of GmFNSII-2^(C),increased flavone synthase abundance and improved the drought resistance.Furthermore,a singlebase mutation in the GmFNSII-2^(C) promoter generated an additional drought response element(CCCCT),which had stronger interaction strength with GmDREB3 and increased its transcriptional activity under drought conditions.The frequency of drought-resistant soybean varieties with Hap 1(Pro:GmFNSII-2^(C))has increased,suggesting that this haplotype may be selected during soybean breeding.In summary,GmFNSII-2^(C) could be used for molecular breeding of drought-tolerant soybean.

Evolution of plant microRNA gene families

MicroRNAs (miRNAs ) 是他们在植物和动物的目标基因的重要 post-transcriptional 管理者。miRNAs 通常长是 20-24 核苷酸。尽管有他们的不平常地小的尺寸， miRNA 基因家庭的进化历史似乎类似于编码 theirprotein 对应物。与在动物染色体的小却丰富的 miRNA 家庭相对照，植物有少数但是更大的 miRNA 基因家庭。植物 miRNA 基因家庭的成员经常是高度类似的，建议经由双人脚踏车基因复制和部分复制事件的最近的扩大。尽管许多 miRNA 基因越过植物种类被保存，一样的基因家庭在不同种类在尺寸和 genomic 组织显著地变化，它可以在目标基因规定引起剂量效果和空间、时间的差别。在这评论，我们在理解植物 miRNA 基因家庭的进化总结当前的进步。

Deep genotyping of the gene GmSNAP facilitates pyramiding resistance to cyst nematode in soybean

Soybean cyst nematode(SCN)is a highly destructive pathogen.The soybean host genome harbors at least two major genes for resistance(rhg1 and Rhg4),as well as a minor locus(SCN3-11).In the present study,a splicing site in GmSNAP11,the potential causal gene of SCN3-11,was identified by comparison of the GmSNAP11 cDNA sequences generated from resistant and susceptible soybean accessions.The sequence information was used to design a codominant CAPS marker,GmSNAP11-2565,which was used to genotype a panel of 209 soybean accessions varying with respect to SCN resistance.Analyses of the effect of the haplotypes formed by GmSNAP11-2565 and another large-effect(nonsynonymous)locus,GmSNAP11-2307,previously identified in GmSNAP11,revealed linkage disequilibrium(P<0.0001)between the two loci,suggesting that GmSNAP11-2565 could be used as a marker for GmSNAP11.GmSNAP11-2565 was accordingly used,along with established markers for GmSNAP18(rhg1)and GmSHMT(Rhg4),to characterize the panel accessions.The mean SCN female index of accessions carrying only the GmSNAP11 allele associated with resistance(20.3%)was higher than that associated with accessions carrying alleles for resistance at both GmSNAP11 and GmSNAP18(12.4%),while the index for accessions carrying alleles for resistance at all of GmSNAP11,GmSNAP18,and GmSHMT was very low(1.9%).Selection on all three markers was effective for maintaining a high level of resistance to SCN race 3.

Identification of novel soybean oil content-related genes using QTL-based collinearity analysis from the collective soybean genome

Soybean is a global principal source of edible plant oil. As more soybean oil-related quantitative trait loci(QTLs) have been located in the collective genome, it is urgent to establish a classification system for these distributed QTLs. A collinear platform may be useful to characterize and identify relationships among QTLs as well as aid in novel gene discovery. In this study, the collinearity MCScan X algorithm and collective soybean genomic information were used to construct collinearity blocks, to which soybean oil-related QTLs were mapped. The results demonstrated that 666 collinearity blocks were detected in the soybean genome across 20 chromosomes, and 521 collinearity relationships existed in 231 of the 242 effective soybean oil-related QTLs. This included 214 inclusion relationships and 307 intersecting relationships. Among them, the collinearity among QTLs that are related to soybean oil content was shown on a maximum of seven chromosomes and minimum of one chromosome, with the majority of QTLs having collinearity on two chromosomes. Using overlapping hotspot regions in the soybean oil QTLs with collinearity, we mined for novel oil content-related genes. Overall, we identified 23 putatively functional genes associated with oil content in soybean and annotated them using a number of annotation databases. Our findings provide a valuable framework for elucidating evolutionary relationships between soybean oil-related QTLs and lay a foundation for functional marker-assisted breeding relating to soybean oil content.

Development and identification of glyphosate-tolerant transgenic soybean via direct selection with glyphosate

Glyphosate-tolerant soybean is the most widely planted genetically modified crop worldwide. However, soybean remains recalcitrant to routine transformation because of the low infection efficiency of Agrobacterium to soybean and lack of useful selectable markers. In this study, several Agrobacterium strains and cell densities were compared by transient expression of the GUS gene. The results showed that Agrobacterium strain Ag10 at cell densities of OD_(600) of 0.6–0.9 yielded the highest infection efficiency in Agrobacterium-mediated soybean cotyledonary node transformation system. Meanwhile, a simple and rapid method was developed for identification of glyphosate tolerance in putative T_0 transgenic plants, consisting of spotting plantlets with 1 μL Roundup~?. The whole cycle of genetic transformation could be shortened to about 3 mon by highly efficient selection with glyphosate during the transformation process and application of the spot assay in putative T_0 transgenic plantlets. The transformation frequency ranged from 2.9 to 5.6%. This study provides an improved protocol for development and identification of glyphosate-tolerant transgenic soybeans.

The identification of presence/absence variants associated with the apparent differences of growth period structures between cultivated and wild soybeans

The cultivated soybean（Glycine max（L.） Merr.） was distinguished from its wild progenitor Glycine soja Sieb.＆ Zucc.in growth period structure,by a shorter vegetative phase（V）,a prolonged reproductive phase（R） and hence a larger R/V ratio.However,the genetic basis of the domestication of soybean from wild materials is unclear.Here,a panel of 123 cultivated and 97 wild accessions were genotyped using a set of 24 presence/absence variants（PAVs） while at the same time the materials were phenotyped with respect to flowering and maturity times at two trial sites located at very different latitudes.The major result of this study showed that variation at PAVs is informative for assessing patterns of genetic diversity in Glycine spp.The genotyping was largely consistent with the taxonomic status,although a few accessions were intermediate between the two major clades identified.Allelic diversity was much higher in the wild germplasm than in the cultivated materials.A significant domestication signal was detected at 11 of the PAVs at 0.01 level.In particular,this study has provided information for revealing the genetic basis of photoperiodism which was a prominent feature for the domestication of soybean.A significant marker-trait association with R/V ratio was detected at 14 of the PAVs,but stripping out population structure reduced this to three.These results will provide markers information for further finding of R/V related genes that can help to understand the domestication process and introgress novel genes in wild soybean to broaden the genetic base of modern soybean cultivars.

Creation of gene-specific rice mutants by AvrXa23-based TALENs

Transcription activator-like effector （TALE） nucleases （TALENs） are increasingly used as a powerful tool for genome edit- ing in a variety of organisms. We have previously cloned the TALE-coding gene avrXa23 from Xanthomonas oryzae pv. oryzae and developed an AvrXa23-based assembly system for designer TALEs or TALENs. Here, we exploit TALENs to induce mutagenesis of the rice ethylene response factor （ERF） transcription factor OsERF922 for testing the gene-editing efficiency of AvrXa23-based TALENs system. A pair of TALENs （T-KJ9/KJ 10） was assembled and their nuclease activities were first confirmed in rice protoplast transient assay. The TALENs-expressing construct pT-KJ9/KJ10 was then used for rice transformation. We observed targeting somatic mutagenesis frequency of 15.0% in positive transgenic rice calli and obtained two mutant plants with nucleotide deletion or insertion at the designer target region. Our work demonstrates that the AvrXa23-based TALENs system can be used for site-specific genome editing in rice.