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1.
Plant Cell Environ ; 47(5): 1656-1667, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38282250

RESUMO

Soybean (Glycine max) is a typical short-day plant, but has been widely cultivated in high-latitude long-day (LD) regions because of the development of early-maturing genotypes which are photoperiod-insensitive. However, some early-maturing varieties exhibit significant responses to maturity under different daylengths but not for flowering, depicting an evident photoperiodic after-effect, a poorly understood mechanism. In this study, we investigated the postflowering responses of 11 early-maturing soybean varieties to various preflowering photoperiodic treatments. We confirmed that preflowering SD conditions greatly promoted maturity and other postflowering developmental stages. Soybean homologs of FLOWERING LOCUS T (FT), including GmFT2a, GmFT3a, GmFT3b and GmFT5a, were highly accumulated in leaves under preflowering SD treatment. More importantly, they maintained a high expression level after flowering even under LD conditions. E1 RNAi and GmFT2a overexpression lines showed extremely early maturity regardless of preflowering SD and LD treatments due to constitutively high levels of floral-promoting GmFT homolog expression throughout their life cycle. Collectively, our data indicate that high and stable expression of floral-promoting GmFT homologs play key roles in the maintenance of photoperiodic induction to promote postflowering reproductive development, which confers early-maturing varieties with appropriate vegetative growth and shortened reproductive growth periods for adaptation to high latitudes.


Assuntos
Glycine max , Fotoperíodo , Glycine max/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Flores/fisiologia , Ritmo Circadiano , Regulação da Expressão Gênica de Plantas
2.
Theor Appl Genet ; 136(12): 245, 2023 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-37962664

RESUMO

KEY MESSAGE: A total of 101 QTNs were found to be associated with soybean flowering time responses to photo-thermal conditions; three candidate genes with non-synonymous substitutions were identified: Glyma.08G302500 (GmHY5), Glyma.08G303900 (GmPIF4c), and Glyma.16G046700 (GmVRN1). The flowering transition is a crucial component of soybean (Glycine max L. Merr.) development. The transition process is regulated by photoperiod, temperature, and their interaction. To examine the genetic architecture associated with temperature- and photo-thermal-mediated regulation of soybean flowering, we here performed a genome-wide association study using a panel of 201 soybean cultivars with maturity groups ranging from MG 000 to VIII. Each cultivar was grown in artificially controlled photoperiod and different seasons in 2017 and 2018 to assess the thermal response (TR) and the interactive photo-thermal response (IPT) of soybean flowering time. The panel contained 96,299 SNPs with minor allele frequencies > 5%; 33, 19, and 49 of these SNPs were significantly associated with only TR, only IPT, and both TR and IPT, respectively. Twenty-one SNPs were located in or near previously reported quantitative trait loci for first-flowering; 16 SNPs were located within 200 kb of the main-effect flowering genes GmFT2a, GmFT2b, GmFT3a, GmFT3b, GmFT5a, GmFT5b, GmCOL2b, GmPIF4b, and GmPIF4c, or near homologs of the known Arabidopsis thaliana flowering genes BBX19, VRN1, TFL1, FUL, AGL19, SPA1, HY5, PFT1, and EDF1. Natural non-synonymous allelic variations were identified in the candidate genes Glyma.08G302500 (GmHY5), Glyma.08G303900 (GmPIF4c), and Glyma.16G046700 (GmVRN1). Cultivars with different haplotypes showed significant variations in TR, IPT, and flowering time in multiple environments. The favorable alleles, candidate genes, and diagnostic SNP markers identified here provide valuable information for future improvement of soybean photo-thermal adaptability, enabling expansion of soybean production regions and improving plant resilience to global climate change.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Glycine max/genética , Estudo de Associação Genômica Ampla , Temperatura , Alelos , Fatores de Transcrição
3.
Theor Appl Genet ; 135(12): 4507-4522, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36422673

RESUMO

KEY MESSAGE: The genetic basis of soybean root system architecture (RSA) and the genetic relationship between shoot and RSA were revealed by integrating data from recombinant inbred population grafting and QTL mapping. Variations in root system architecture (RSA) affect the functions of roots and thus play vital roles in plant adaptations and agricultural productivity. The aim of this study was to unravel the genetic relationship between RSA traits and shoot-related traits in soybean. This study characterized RSA variability at seedling stage in a recombinant inbred population, derived from a cross between cultivated soybean C08 and wild soybean W05, and performed high-resolution quantitative trait locus (QTL) mapping. In total, 34 and 41 QTLs were detected for RSA-related and shoot-related traits, respectively, constituting eight QTL clusters. Significant QTL correspondence was found between shoot biomass and RSA-related traits, consistent with significant correlations between these phenotypes. RSA-related QTLs also overlapped with selection regions in the genome, suggesting the cultivar RSA could be a partial consequence of domestication. Using reciprocal grafting, we confirmed that shoot-derived signals affected root development and the effects were controlled by multiple loci. Meanwhile, RSA-related QTLs were found to co-localize with four soybean flowering-time loci. Consistent with the phenotypes of the parental lines of our RI population, diminishing the function of flowering controlling E1 family through RNA interference (RNAi) led to reduced root growth. This implies that the flowering time-related genes within the RSA-related QTLs are actually contributing to RSA. To conclude, this study identified the QTLs that determine RSA through controlling root growth indirectly via regulating shoot functions, and discovered superior alleles from wild soybean that could be used to improve the root structure in existing soybean cultivars.


Assuntos
Glycine max , Locos de Características Quantitativas , Glycine max/genética , Raízes de Plantas/genética , Mapeamento Cromossômico , Fenótipo
4.
Int J Mol Sci ; 23(17)2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36077363

RESUMO

Pseudo-response regulator (PRR) family members serve as key components of the core clock of the circadian clock, and play important roles in photoperiodic flowering, stress tolerance, growth, and the development of plants. In this study, 14 soybean PRR genes were identified, and classified into three groups according to phylogenetic analysis and structural characteristics. Real-time quantitative PCR analysis revealed that 13 GmPRRs exhibited obvious rhythmic expression under long-day (LD) and short-day (SD) conditions, and the expression of 12 GmPRRs was higher under LD in leaves. To evaluate the effects of natural variations in GmPRR alleles on soybean adaptation, we examined the sequences of GmPRRs among 207 varieties collected across China and the US, investigated the flowering phenotypes in six environments, and analyzed the geographical distributions of the major haplotypes. The results showed that a majority of non-synonymous mutations in the coding region were associated with flowering time, and we found that the nonsense mutations resulting in deletion of the CCT domain were related to early flowering. Haplotype analysis demonstrated that the haplotypes associated with early flowering were mostly distributed in Northeast China, while the haplotypes associated with late flowering were mostly cultivated in the lower latitudes of China. Our study of PRR family genes in soybean provides not only an important guide for characterizing the circadian clock-controlled flowering pathway but also a theoretical basis and opportunities to breed varieties with adaptation to specific regions and farming systems.


Assuntos
Regulação da Expressão Gênica de Plantas , Glycine max , Flores , Genômica , Fotoperíodo , Filogenia , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Glycine max/metabolismo
5.
Plant Cell Environ ; 44(8): 2551-2564, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34050544

RESUMO

Soybean (Glycine max), a typical short-day plant (SDP) domesticated in temperate regions, has expanded to high latitudes where daylengths are long from soybean emergence to bloom, but rapidly decrease from seed filling to maturity. Cotyledons are well known as the major storage organs in seeds, but it is unclear whether developing cotyledons store flowering substances at filling stage in SD for upcoming seedlings, or instead respond to photoperiod for floral induction after emergence of matured seeds in long-day (LD). Here, we report that cotyledons accelerate flowering of early-maturing varieties not resulting from stored floral stimuli but by perceiving photoperiod after emergence. We found that light signal is indispensable to activate cotyledons for floral induction, and flowering promoting gene GmFT2a is required for cotyledon-dependent floral induction via upregulation of floral identity gene GmAP1. Interestingly, cotyledons are competent to support the entire life cycle of a cotyledon-only plant to produce seeds, underlying a new photoperiod study system in soybean and other dicots. Taken together, these results demonstrate a substantial role for cotyledons in flowering process, whereby we propose a 'cotyledon-based self-reliance' model highlighting floral induction from emergence as a key ecological adaptation for rapid flowering of SDPs grown in LD environments at high latitudes.


Assuntos
Adaptação Fisiológica , Cotilédone/fisiologia , Glycine max/fisiologia , China , Flores/fisiologia , Regulação da Expressão Gênica de Plantas , Luz , Fotoperíodo , Plantas Geneticamente Modificadas , Proteínas de Soja/genética
6.
Plant Biotechnol J ; 18(9): 1869-1881, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-31981443

RESUMO

Flowering time is a critical determinant of the geographic distribution and regional adaptability of soybean (Glycine max) and is strongly regulated by photoperiod and temperature. In this study, quantitative trait locus (QTL) mapping and subsequent candidate gene analysis revealed that GmPRR37, encoding a pseudo-response regulator protein, is responsible for the major QTL qFT12-2, which was identified from a population of 308 recombinant inbred lines (RILs) derived from a cross between a very late-flowering soybean cultivar, 'Zigongdongdou (ZGDD)', and an extremely early-flowering cultivar, 'Heihe27 (HH27)', in multiple environments. Comparative analysis of parental sequencing data confirmed that HH27 contains a non-sense mutation that causes the loss of the CCT domain in the GmPRR37 protein. CRISPR/Cas9-induced Gmprr37-ZGDD mutants in soybean exhibited early flowering under natural long-day (NLD) conditions. Overexpression of GmPRR37 significantly delayed the flowering of transgenic soybean plants compared with wild-type under long photoperiod conditions. In addition, both the knockout and overexpression of GmPRR37 in soybean showed no significant phenotypic alterations in flowering time under short-day (SD) conditions. Furthermore, GmPRR37 down-regulated the expression of the flowering-promoting FT homologues GmFT2a and GmFT5a, and up-regulated flowering-inhibiting FT homologue GmFT1a expression under long-day (LD) conditions. We analysed haplotypes of GmPRR37 among 180 cultivars collected across China and found natural Gmprr37 mutants flower earlier and enable soybean to be cultivated at higher latitudes. This study demonstrates that GmPRR37 controls soybean photoperiodic flowering and provides opportunities to breed optimized cultivars with adaptation to specific regions and farming systems.


Assuntos
Glycine max , Fotoperíodo , Sistemas CRISPR-Cas/genética , China , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Mutação/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Glycine max/genética , Glycine max/metabolismo
7.
Plant Biotechnol J ; 18(1): 298-309, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31240772

RESUMO

Flowering time is a key agronomic trait that directly influences the successful adaptation of soybean (Glycine max) to diverse latitudes and farming systems. GmFT2a and GmFT5a have been extensively identified as flowering activators and integrators in soybean. Here, we identified two quantitative trait loci (QTLs) regions harbouring GmFT2a and GmFT5a, respectively, associated with different genetic effects on flowering under different photoperiods. We analysed the flowering time of transgenic plants overexpressing GmFT2a or GmFT5a, ft2a mutants, ft5a mutants and ft2aft5a double mutants under long-day (LD) and short-day (SD) conditions. We confirmed that GmFT2a and GmFT5a are not redundant, they collectively regulate flowering time, and the effect of GmFT2a is more prominent than that of GmFT5a under SD conditions whereas GmFT5a has more significant effects than GmFT2a under LD conditions. GmFT5a, not GmFT2a, was essential for soybean to adapt to high latitude regions. The ft2aft5a double mutants showed late flowering by about 31.3 days under SD conditions and produced significantly increased numbers of pods and seeds per plant compared to the wild type. We speculate that these mutants may have enormous yield potential for the tropics. In addition, we examined the sequences of these two loci in 202 soybean accessions and investigated the flowering phenotypes, geographical distributions and maturity groups within major haplotypes. These results will contribute to soybean breeding and regional adaptability.


Assuntos
Sistemas CRISPR-Cas , Flores/crescimento & desenvolvimento , Glycine max/genética , Locos de Características Quantitativas , Adaptação Biológica/genética , Mutagênese , Fotoperíodo , Proteínas de Plantas/genética
8.
Plant Cell Environ ; 43(4): 934-944, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31981430

RESUMO

Day length has an important influence on flowering and growth habit in many plant species. In crops such as soybean, photoperiod sensitivity determines the geographical range over which a given cultivar can grow and flower. The soybean genome contains ~10 genes homologous to FT, a central regulator of flowering from Arabidopsis thaliana. However, the precise roles of these soybean FTs are not clearly. Here we show that one such gene, GmFT2b, promotes flowering under long-days (LDs). Overexpression of GmFT2b upregulates expression of flowering-related genes which are important in regulating flowering time. We propose a 'weight' model for soybean flowering under short-day (SD) and LD conditions. Furthermore, we examine GmFT2b sequences in 195 soybean cultivars, as well as flowering phenotypes, geographical distributions and maturity groups. We found that Hap3, a major GmFT2b haplotype, is associated with significantly earlier flowering at higher latitudes. We anticipate our assay to provide important resources for the genetic improvement of soybean, including new germplasm for soybean breeding, and also increase our understanding of functional diversity in the soybean FT gene family.


Assuntos
Glycine max/fisiologia , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Adaptação Fisiológica/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiologia , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Clonagem Molecular , Flores/crescimento & desenvolvimento , Edição de Genes , Regulação da Expressão Gênica de Plantas/genética , Variação Genética/genética , Variação Genética/fisiologia , Geografia , Fotoperíodo , Proteínas de Plantas/fisiologia , Glycine max/genética , Glycine max/crescimento & desenvolvimento , Fatores de Transcrição/fisiologia , Transcriptoma
9.
BMC Genomics ; 20(1): 230, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30894121

RESUMO

BACKGROUND: Flowering time and maturity are among the most important adaptive traits in soybean (Glycine max (L.) Merill). Flowering Locus T (FT) family genes function as key flowering integrators, with flowering-promoting members GmFT2a/GmFT5a and flowering-inhibiting members GmFT4/GmFT1a antagonistically regulating vegetative and reproductive growth. However, to date, the relations between natural variations of FT family genes and the diversity of flowering time and maturity in soybean are not clear. Therefore, we conducted this study to discover natural variations in FT family genes in association with flowering time and maturity. RESULTS: Ten FT family genes, GmFT1a, GmFT1b, GmFT2a, GmFT2b, GmFT3a, GmFT3b, GmFT4, GmFT5a, GmFT5b and GmFT6, were cloned and sequenced in the 127 varieties evenly covering all 14 known maturity groups (MG0000-MGX). They were diversified at the genome sequence polymorphism level. GmFT3b and GmFT5b might have experienced breeding selection in the process of soybean domestication and breeding. Haplotype analysis showed that a total of 17 haplotypes had correlative relationships with flowering time and maturity among the 10 FT genes, namely, 1a-H3, 1b-H1, 1b-H6, 1b-H7, 2a-H1, 2a-H3, 2a-H4, 2a-H9, 2b-H3, 2b-H4, 2b-H6, 2b-H7, 3b-H4, 5a-H1, 5a-H2, 5a-H4 and 5b-H1. Based on the association analysis, 38 polymorphic sites had a significant association with flowering time at the level of p < 0.01. CONCLUSIONS: Some natural variations exist within the 10 FT family genes, which might be involved in soybean adaptation to different environments and have an influence on diverse flowering time and maturity. This study will facilitate the understanding of the roles of FTs in flowering and maturity.


Assuntos
Variação Genética , Glycine max/crescimento & desenvolvimento , Glycine max/genética , Proteínas de Plantas/genética , Haplótipos , Polimorfismo Genético , Fatores de Tempo
10.
Int J Mol Sci ; 20(23)2019 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-31775326

RESUMO

Soybean is an excellent source of vegetable protein and edible oil. Understanding the genetic basis of protein and oil content will improve the breeding programs for soybean. Linkage analysis and genome-wide association study (GWAS) tools were combined to detect quantitative trait loci (QTL) that are associated with protein and oil content in soybean. Three hundred and eight recombinant inbred lines (RILs) containing 3454 single nucleotide polymorphism (SNP) markers and 200 soybean accessions, including 94,462 SNPs and indels, were applied to identify QTL intervals and significant SNP loci. Intervals on chromosomes 1, 15, and 20 were correlated with both traits, and QTL qPro15-1, qPro20-1, and qOil5-1 reproducibly correlated with large phenotypic variations. SNP loci on chromosome 20 that overlapped with qPro20-1 were reproducibly connected to both traits by GWAS (p < 10-4). Twenty-five candidate genes with putative roles in protein and/or oil metabolisms within two regions (qPro15-1, qPro20-1) were identified, and eight of these genes showed differential expressions in parent lines during late reproductive growth stages, consistent with a role in controlling protein and oil content. The new well-defined QTL should significantly improve molecular breeding programs, and the identified candidate genes may help elucidate the mechanisms of protein and oil biosynthesis.


Assuntos
Ligação Genética , Estudo de Associação Genômica Ampla , Glycine max/genética , Óleos de Plantas/metabolismo , Locos de Características Quantitativas , Sementes/genética , Proteínas de Soja/genética , Mapeamento Cromossômico , Cromossomos de Plantas , Genoma de Planta , Desequilíbrio de Ligação , Fenótipo , Polimorfismo de Nucleotídeo Único , Sementes/metabolismo , Proteínas de Soja/metabolismo , Glycine max/metabolismo
11.
Plant Biotechnol J ; 16(1): 176-185, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28509421

RESUMO

Flowering is an indication of the transition from vegetative growth to reproductive growth and has considerable effects on the life cycle of soya bean (Glycine max). In this study, we employed the CRISPR/Cas9 system to specifically induce targeted mutagenesis of GmFT2a, an integrator in the photoperiod flowering pathway in soya bean. The soya bean cultivar Jack was transformed with three sgRNA/Cas9 vectors targeting different sites of endogenous GmFT2a via Agrobacterium tumefaciens-mediated transformation. Site-directed mutations were observed at all targeted sites by DNA sequencing analysis. T1-generation soya bean plants homozygous for null alleles of GmFT2a frameshift mutated by a 1-bp insertion or short deletion exhibited late flowering under natural conditions (summer) in Beijing, China (N39°58', E116°20'). We also found that the targeted mutagenesis was stably heritable in the following T2 generation, and the homozygous GmFT2a mutants exhibited late flowering under both long-day and short-day conditions. We identified some 'transgene-clean' soya bean plants that were homozygous for null alleles of endogenous GmFT2a and without any transgenic element from the T1 and T2 generations. These 'transgene-clean' mutants of GmFT2a may provide materials for more in-depth research of GmFT2a functions and the molecular mechanism of photoperiod responses in soya bean. They will also contribute to soya bean breeding and regional introduction.


Assuntos
Sistemas CRISPR-Cas/genética , Flores/metabolismo , Flores/fisiologia , Glycine max/metabolismo , Glycine max/fisiologia , Sistemas CRISPR-Cas/fisiologia , Flores/genética , Edição de Genes , Genoma de Planta/genética , Mutagênese/genética , Mutagênese/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Glycine max/genética
12.
New Phytol ; 217(3): 1335-1345, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29120038

RESUMO

Soybean flowering and maturation are strictly regulated by photoperiod. Photoperiod-sensitive soybean varieties can undergo flowering reversion when switched from short-day (SD) to long-day (LD) conditions, suggesting the presence of a 'floral-inhibitor' under LD conditions. We combined gene expression profiling with a study of transgenic plants and confirmed that GmFT1a, soybean Flowering Locus T (FT) homolog, is a floral inhibitor. GmFT1a is expressed specifically in leaves, similar to the flowering-promoting FT homologs GmFT2a/5a. However, in Zigongdongdou (ZGDD), a model variety for studying flowering reversion, GmFT1a expression was induced by LD but inhibited by SD conditions. This was unexpected, as it is the complete opposite of the expression of flowering promoters GmFT2a/5a. Moreover, the key soybean maturity gene E1 may up-regulate GmFT1a expression. It is also notable that GmFT1a expression was conspicuously high in late-flowering varieties. Transgenic overexpression of GmFT1a delayed flowering and maturation in soybean, confirming that GmFT1a functions as a flowering inhibitor. This discovery highlights the complex impacts of the functional diversification of the FT gene family in soybean, and implies that antagonism between flowering-inhibiting and flowering-promoting FT homologs in this highly photoperiod-sensitive plant may specify vegetative vs reproductive development.


Assuntos
Flores/fisiologia , Glycine max/genética , Proteínas de Plantas/genética , Homologia de Sequência de Aminoácidos , Flores/genética , Regulação da Expressão Gênica de Plantas , Haplótipos/genética , Modelos Biológicos , Fenótipo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Transcriptoma/genética
13.
Int J Mol Sci ; 19(10)2018 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-30301169

RESUMO

As a genetically modified crop, transgenic soybean occupies the largest global scale with its food, nutritional, industrial, and pharmaceutical uses.Efficient transformation is a key factor for the improvement of genetically modified soybean. At present, the Agrobacterium-mediated method is primarily used for soybean transformation, but the efficiency of this method is still relatively low (below 5%) compared with rice (above 90%). In this study, we examined the influence of l-glutamine and/or l-asparagine on Agrobacterium-mediated transformation in soybean and explored the probable role in the process of Agrobacterium-mediated transformation. The results showed that when the amino acids l-glutamine and l-asparagine were added separately or together to the culture medium, the shoot induction frequency, elongation rate, and transformation frequency were improved. The combined effects of l-glutamine and l-asparagine were better than those of l-glutamine and l-asparagine alone. The 50 mg/L l-glutamine and 50 mg/L l-asparagine together can enhance the transformation frequency of soybean by attenuating the expression level of GmPRs (GmPR1, GmPR4, GmPR5, and GmPR10) and suppression of the plant defense response. The transgene was successfully transmitted to the T1 generation. This study will be useful in genetic engineering of soybean.


Assuntos
Agrobacterium tumefaciens/genética , Asparagina/farmacologia , Meios de Cultura/farmacologia , Glutamina/farmacologia , Glycine max/genética , Transformação Genética/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Engenharia Genética , Raízes de Plantas/efeitos dos fármacos , Brotos de Planta/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética , Transgenes/genética
14.
Int J Mol Sci ; 19(12)2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30513774

RESUMO

At present, the application of CRISPR/Cas9 in soybean (Glycine max (L.) Merr.) has been mainly focused on knocking out target genes, and most site-directed mutagenesis has occurred at single cleavage sites and resulted in short deletions and/or insertions. However, the use of multiple guide RNAs for complex genome editing, especially the deletion of large DNA fragments in soybean, has not been systematically explored. In this study, we employed CRISPR/Cas9 technology to specifically induce targeted deletions of DNA fragments in GmFT2a (Glyma16g26660) and GmFT5a (Glyma16g04830) in soybean using a dual-sgRNA/Cas9 design. We achieved a deletion frequency of 15.6% for target fragments ranging from 599 to 1618 bp in GmFT2a. We also achieved deletion frequencies of 12.1% for target fragments exceeding 4.5 kb in GmFT2a and 15.8% for target fragments ranging from 1069 to 1161 bp in GmFT5a. In addition, we demonstrated that these CRISPR/Cas9-induced large fragment deletions can be inherited. The T2 'transgene-free' homozygous ft2a mutants with a 1618 bp deletion exhibited the late-flowering phenotype. In this study, we developed an efficient system for deleting large fragments in soybean using CRISPR/Cas9; this system could benefit future research on gene function and improve agriculture via chromosome engineering or customized genetic breeding in soybean.


Assuntos
Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas/genética , Genoma de Planta , Glycine max/genética , Sequência de Bases , Genes de Plantas , Homozigoto , Padrões de Herança/genética , Mutagênese Sítio-Dirigida , Mutação/genética , Taxa de Mutação , Fenótipo , Deleção de Sequência
15.
World J Microbiol Biotechnol ; 34(8): 113, 2018 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-29987404

RESUMO

Genetically modified (GM) crops have brought various economic benefits but may also have adversely affected soil microorganisms. To examine whether transgenic high-methionine soybean ZD91 alters the bacterial community structure in the rhizosphere, we performed a 2-year follow-up study using the transgenic high-methionine soybean cultivar ZD91 and wild type cultivar ZD. The community composition and the relative abundance of bacteria in rhizosphere soil were determined by sequencing of the 16S rRNA amplicon. Our results indicated that transgenic soybean ZD91 had no significantly effects on rhizosphere bacterial communities. Instead, the plant growth stage and year appeared to have a stronger effect on bacterial communities. Our findings therefore provided reliable scientific evidence for potential commercial cultivation of cultivar ZD91.


Assuntos
Bactérias/classificação , Glycine max/microbiologia , Metionina/metabolismo , Consórcios Microbianos , Plantas Geneticamente Modificadas/microbiologia , Rizosfera , Microbiologia do Solo , Bactérias/genética , Biodiversidade , DNA Bacteriano/análise , Seguimentos , Consórcios Microbianos/genética , Filogenia , Raízes de Plantas/microbiologia , Plantas Geneticamente Modificadas/metabolismo , RNA Ribossômico 16S/genética , Análise de Sequência , Solo , Glycine max/crescimento & desenvolvimento
16.
BMC Genomics ; 18(1): 415, 2017 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-28549456

RESUMO

BACKGROUND: Soybean (Glycine max (L.) Merr.) is a short day plant. Its flowering and maturity time are controlled by genetic and environmental factors, as well the interaction between the two factors. Previous studies have shown that both genetic and environmental factors, mainly photoperiod and temperature, control flowering time of soybean. Additionally, these studies have reported gene × gene and gene × environment interactions on flowering time. However, the effects of quantitative trait loci (QTL) in response to photoperiod and temperature have not been well evaluated. The objectives of the current study were to identify the effects of loci associated with flowering time under different photo-thermal conditions and to understand the effects of interaction between loci and environment on soybean flowering. METHODS: Different photoperiod and temperature combinations were obtained by adjusting sowing dates (spring sowing and summer sowing) or day-length (12 h, 16 h). Association mapping was performed on 91 soybean cultivars from different maturity groups (MG000-VIII) using 172 SSR markers and 5107 SNPs from the Illumina SoySNP6K iSelectBeadChip. The effects of the interaction between QTL and environments on flowering time were also analysed using the QTXNetwork. RESULTS: Large-effect loci were detected on Gm 11, Gm 16 and Gm 20 as in previous reports. Most loci associated with flowering time are sensitive to photo-thermal conditions. Number of loci associated with flowering time was more under the long day (LD) than under the short day (SD) condition. The variation of flowering time among the soybean cultivars mostly resulted from the epistasis × environment and additive × environment interactions. Among the three candidate loci, i.e. Gm04_4497001 (near GmCOL3a), Gm16_30766209 (near GmFT2a and GmFT2b) and Gm19_47514601 (E3 or GmPhyA3), the Gm04_4497001 may be the key locus interacting with other loci for controlling soybean flowering time. CONCLUSION: The effects of loci associated with the flowering time of soybean were dependent upon the photo-thermal conditions. This study facilitates the understanding of the genetic mechanism of soybean flowering and molecular breeding for the improvement of soybean adaptability to specific and/or broad regions.


Assuntos
Mapeamento Cromossômico , Flores/crescimento & desenvolvimento , Interação Gene-Ambiente , Glycine max/crescimento & desenvolvimento , Glycine max/genética , Fotoperíodo , Temperatura , Genótipo , Desequilíbrio de Ligação , Polimorfismo de Nucleotídeo Único
17.
Microbiology (Reading) ; 163(2): 197-206, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27995867

RESUMO

Pantoea agglomerans YS19 is a preponderant endophytic bacterium isolated from rice. It is characterized by the formation of symplasmata, a type of multicellular aggregate structure, contributing to a strong stress resistance and specific adaptation of YS19 in endophyte-host associations. Indole is an important signal molecule in intra- or interspecies relationships, regulating a variety of bacterial behaviours such as cell aggregation and stress resistance; however, the regulatory mechanism remains an ongoing area of investigation. This study selected YS19 as a model strain to construct a mutant library, utilizing the mTn5 transposon mutagenesis method, thus obtaining a positive mutant with an indole-inhibited mutation gene. Via thermal asymmetric interlaced PCR, the mutational site was identified as the gene of pcnB, which encodes the poly(A) polymerase I to catalyse the polyadenylation of RNAs. The full length of the pcnB sequence was 1332 bp, and phylogenetic analysis revealed that pcnB is extremely conserved among strains of P. agglomerans. The expression of the gene was significantly inhibited (by 36.6 % as detected via quantitative PCR) by indole (0.5 mM). Many physiological behaviours of YS19 were affected by this mutation: the cell decay rate in the post-stationary growth phase was promoted, symplasmata formation and motility were inhibited in the late stationary growth phase and the colonization ability and growth-promoting effect of YS19 on the host plant were also inhibited. This study discusses the indole regulatory pathways from the point of RNA post-transcriptional modification, thus enriching our knowledge of polyadenylation and expanding current research ideas of indole regulation.


Assuntos
Aderência Bacteriana/fisiologia , Indóis/metabolismo , Pantoea/metabolismo , Poliadenilação/fisiologia , Polinucleotídeo Adenililtransferase/metabolismo , Sequência de Aminoácidos , Aderência Bacteriana/genética , Pantoea/genética , Poliadenilação/genética , Polinucleotídeo Adenililtransferase/genética
20.
Plant Cell Rep ; 34(1): 121-32, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25326369

RESUMO

KEY MESSAGE: A FRUITFULL homolog GmFULa was cloned and found to play roles in the flowering and maturation of soybean. Soybean varieties exhibit great diversity in terms of flowering and maturation due to differences in their photoperiodic responses. The underlying mechanism remains unclear despite the fact that some upstream flowering genes have been studied. FRUITFULL (FUL) genes are one group of downstream flowering genes known to have major roles in reproductive transition, floral meristem identity, and floral organ identity. However, FUL homologs and their functions are poorly understood in soybean. Here, a soybean FUL homolog was cloned from the late-maturing photoperiod-sensitive soybean variety Zigongdongdou (ZGDD) and designated GmFULa. In ZGDD, GmFULa exhibited a terminal-preferential expression pattern, with higher expression in the root and shoot apices than in the middle parts. Diurnal rhythm analysis revealed that photoperiod regulates the GmFULa expression level but does not alter its diurnal rhythm. ZGDD was maintained under different photoperiod conditions (long day, LD; short day, SD; LD after 13 short days, SD13-LD) to assess GmFULa expression in newly expanded leaves and in the shoot apex. From this analysis, GmFULa expression was detected in the floral meristem, floral organs and their primordia; trifoliate leaves; and the inflorescence meristem, with the expression levels induced by SD and inhibited by LD. GmFULa expression was also associated with maturity in seven soybean varieties with different photoperiod sensitivities. Therefore, photoperiod conditions affect the expression level of GmFULa but not its diurnal rhythm. The gene plays pleiotropic roles in reproductive transition, flowering, and leaf development and is associated with maturity in soybean.


Assuntos
Flores/genética , Glycine max/genética , Proteínas de Domínio MADS/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Hibridização In Situ , Proteínas de Domínio MADS/classificação , Meristema/genética , Meristema/crescimento & desenvolvimento , Dados de Sequência Molecular , Fotoperíodo , Filogenia , Proteínas de Plantas/classificação , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Glycine max/crescimento & desenvolvimento
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