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1.
Hai Ping Hong Hongyi Zhang Paula Olhoft Steve Hill Hunt Wiley Effie Toren Helke Hillebrand Todd Jones Ming Cheng 《In vitro cellular & developmental biology. Plant》2007,43(6):558-568
A regeneration and transformation system has been developed using organogenic calluses derived from soybean axillary nodes
as the starting explants. Leaf-node or cotyledonary-node explants were prepared from 7 to 8-d-old seedlings. Callus was induced
on medium containing either Murashige and Skoog (MS) salts or modified Finer and Nagasawa (FNL) salts and B5 vitamins with
various concentrations of benzylamino purine (BA) and thidiazuron (TDZ). The combination of BA and TDZ had a synergistic effect
on callus induction. Shoot differentiation from the callus occurred once the callus was transferred to medium containing a
low concentration of BA. Subsequently, shoots were elongated on medium containing indole-3-acetic acid (IAA), zeatin riboside,
and gibberellic acid (GA). Plant regeneration from callus occurred 90 ∼ 120 d after the callus was cultured on shoot induction
medium. Both the primary callus and the proliferated callus were used as explants for Agrobacterium-mediated transformation. The calluses were inoculated with A. tumefaciens harboring a binary vector with the bar gene as the selectable marker gene and the gusINT gene for GUS expression. Usually 60–100% of the callus showed transient GUS expression 5 d after inoculation. Infected
calluses were then selected on media amended with various concentrations of glufosinate. Transgenic soybean plants have been
regenerated and established in the greenhouse. GUS expression was exhibited in various tissues and plant organs, including
leaf, stem, and roots. Southern and T1 plant segregation analysis of transgenic events showed that transgenes were integrated into the soybean genome with a copy
number ranging from 1–5 copies. 相似文献
2.
Modern genetic analysis and manipulation of soybean (Glycine max) depend heavily on an efficient and dependable transformation process, especially in public genotypes from which expressed sequence tag (EST), bacterial artificial chromosome and microarray data have been derived. Williams 82 is the subject of EST and functional genomics analyses. However, it has not previously been transformed successfully using either somatic embryogenesis-based or cotyledonary-node transformation methods, the two predominant soybean transformation systems. An advance has recently been made in using antioxidants to enhance Agrobacterium infection of soybean. Nonetheless, an undesirable effect of using these antioxidants is the compromised recovery of transgenic soybean when combined with the use of the herbicide glufosinate as a selective agent. Therefore, we optimized both Agrobacterium infection and glufosinate selection in the presence of l-cysteine for Williams 82. We have recovered transgenic lines of this genotype with an enhanced transformation efficiency using this herbicide selection system.Abbreviations
DTT
Dithiothreitol
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EST
Expressed sequence tag
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GUS
-Glucuronidase
Communicated by P. Ozias-Akins 相似文献
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AR Torres G Kaschuk GP Saridakis M Hungria 《World journal of microbiology & biotechnology》2012,28(4):1831-1835
Brazil has succeeded in sustaining production of soybean [Glycine max (L.) Merrill] by relying mainly on symbiotic N2 fixation, thanks to the selection and use in inoculants of very effective strains of Bradyrhizobium japonicum and Bradyrhizobium elkanii. It is desirable that rhizobial strains used in inoculants have stable genetic and physiological traits, but experience confirms
that rhizobial strains nodulating soybean often lose competitiveness in the field. In this study, soybean cultivar BR 16 was
single-inoculated with four B. japonicum strains (CIAT 88, CIAT 89, CIAT 104 and CIAT 105) under aseptic conditions. Forty colonies were isolated from nodules produced
by each strain. The progenitor strains, the isolates and four other commercially recommended strains were applied separately
to the same cultivar under controlled greenhouse conditions. We observed significant variability in nodulation, shoot dry
weight, shoot total N, nodule efficiency (total N mass over nodule mass) and BOX-PCR fingerprinting profiles between variant
and progenitor strains. Some variant strains resulted in significantly larger responses in terms of shoot total N, dry weight
and nodule efficiency, when compared to their progenitor strain. These results highlight the need for intermittent evaluation
of stock bacterial cultures to guarantee effective symbiosis after inoculation. Most importantly, it indicates that it is
possible to improve symbiotic effectiveness by screening rhizobial strains for higher N2 fixation capacity within the natural variability that can be found within each progenitor strain. 相似文献
5.
The objective of this article is to study the effect of 5-aminolevulinic acid (ALA) and enhanced chlorophyll content, antioxidative
enzymes and photosynthesis rate by foliar application of ALA. We evaluated three concentrations (control-distilled water,
T1-50 mg l−1, T2-150 mg l−1, T3-250 mg l−1) of ALA and seven cultivars, “Sanchidaye” (Sa-1), “Lichuandasuomian” (Li-1), “Aijiaohuang” (Ai-1), “Qingyou” No. 4 (Qi-1),
“Aikang” No. 5 (Ak-1), “Hanxiao” (Ha-1) and “Shulv” (Sl-1). “Ak-1” showed strongest response of POD (peroxidase) enzyme activity
(0.4 U g−1 min−1) in 250 mg l−1 ALA solution. The highest CAT (catalase) activity (0.8 U g−1 min−1) after administration of 250 mg l−1 ALA was observed in “Li-1”. Meanwhile, highest (1.42 mg l−1) total chlorophyll content was also observed in “Ak-1”, when leaves were treated in 50 mg l−1 ALA, “Li-1” and “Ai-1” showed strongest response of specific activity of superoxide dismutase (SOD) in 50 mg l−1 and 50 mg l−1 ALA. Two hundred and fifty milligram per milliliter of ALA-treatment significantly improved the net photosynthetic rate. 相似文献
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Yanbo Cheng Qibin Ma Hailong Ren Qiuju Xia Enliang Song Zhiyuan Tan Shuxian Li Gengyun Zhang Hai Nian 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2017,130(5):1041-1051
Key message
Using a combination of phenotypic screening, genetic and statistical analyses, and high-throughput genome-wide sequencing, we have finely mapped a dominant Phytophthora resistance gene in soybean cultivar Wayao.Abstract
Phytophthora root rot (PRR) caused by Phytophthora sojae is one of the most important soil-borne diseases in many soybean-production regions in the world. Identification of resistant gene(s) and incorporating them into elite varieties are an effective way for breeding to prevent soybean from being harmed by this disease. Two soybean populations of 191 F2 individuals and 196 F7:8 recombinant inbred lines (RILs) were developed to map Rps gene by crossing a susceptible cultivar Huachun 2 with the resistant cultivar Wayao. Genetic analysis of the F2 population indicated that PRR resistance in Wayao was controlled by a single dominant gene, temporarily named RpsWY, which was mapped on chromosome 3. A high-density genetic linkage bin map was constructed using 3469 recombination bins of the RILs to explore the candidate genes by the high-throughput genome-wide sequencing. The results of genotypic analysis showed that the RpsWY gene was located in bin 401 between 4466230 and 4502773 bp on chromosome 3 through line 71 and 100 of the RILs. Four predicted genes (Glyma03g04350, Glyma03g04360, Glyma03g04370, and Glyma03g04380) were found at the narrowed region of 36.5 kb in bin 401. These results suggest that the high-throughput genome-wide resequencing is an effective method to fine map PRR candidate genes.7.
8.
Zhong-feng Li Yong Guo Lin Ou Huilong Hong Jun Wang Zhang-xiong Liu Bingfu Guo Lijuan Zhang Lijuan Qiu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2018,131(5):1001-1016
Key message
GmDW1 encodes an ent-kaurene synthase (KS) acting at the early step of the biosynthesis pathway for gibberellins (GAs) and regulates the development of plant height in soybean.Abstract
Plant height is an important component of plant architecture, and significantly affects crop breeding practices and yield. Here, we report the characterization of an EMS-induced dwarf mutant (dw) of the soybean cultivar Zhongpin 661 (ZDD23893). The dw mutant displayed reduced plant height and shortened internodes, both of which were mainly attributed to the longitudinally decreased cell length. The bioactive GA1 (gibberellin A1) and GA4 (gibberellin A4) were not detectable in the stem of dw, and the dwarf phenotype could be rescued by treatment with exogenous GA3. Genetic analysis showed that the dwarf trait of dw was controlled by a recessive nuclear gene. By combining linkage analysis and mapping-by-sequencing, we mapped the GmDW1 gene to an approximately 460-kb region on chromosome (Chr.) 8, containing 36 annotated genes in the reference Willliams 82 genome. Of these genes, we identified two nonsynonymous single nucleotide polymorphisms (SNPs) that are present in the encoding regions of Gmdw1 and Glyma.08G165100 in dw, respectively. However, only the SNP mutation (T>A) at nucleotide 1224 in Gmdw1 cosegregated with the dwarf phenotype. GmDW1 encodes an ent-kaurene synthase, and was expressed in various tissues including root, stem, and leaf. Further phenotypic analysis of the allelic variations in soybean accessions strongly indicated that GmDW1 is responsible for the dwarf phenotype in dw. Our results provide important information for improving our understanding of the genetics of soybean plant height and crop breeding.9.
Although light is the ultimate substrate in photosynthesis, strong light can also be harmful and lead to photoinhibition. The DEG proteases play important roles in the degradation of misfolded and damaged proteins. In this study, two photoinhibition-related genes from soybean [Glycine max (L.) Merr.], GmDeg1 and GmDeg2, were cloned. Bioinformatics analysis indicated that these two proteases both contain a PDZ domain and are serine proteases. The expression levels of GmDeg1 and GmDeg2 increased significantly after 12 h of photooxidation treatment, indicating that GmDeg1 and GmDeg2 might play protective roles under strong light conditions. In in vitro proteolytic degradation assays, recombinant GmDeg1 and GmDeg2 demonstrated biological activities at temperatures ranging from 20°C to 60°C and at pH 5.0 to 8.0. By contrast, the proteases showed no proteolytic effect in the presence of a serine protease inhibitor. Taken together, these results provided strong evidence that GmDeg1 and GmDeg2 are serine proteases that could degrade the model substrate in vitro, indicating that they might degrade damaged D1 protein and other mis-folded proteins in vivo. Furthermore, GmDeg1 and GmDeg2 were transformed into Arabidopsis thaliana to obtain transgenic plants. Leaves from the transgenic and wild-type plants were subjected to strong light conditions in vitro, and the PSII photochemical efficiency (Fv/Fm) was measured. The Fv/Fm of the transgenic plants was significantly higher than that of the wild-type plants at most time points. These results imply that GmDeg1 and GmDeg2 would have similar functions to Arabidopsis AtDeg1, thus accelerating the recovery of PSII photochemical efficiency. 相似文献
10.
To investigate the biocontrol effectiveness of the antibiotic producing bacterium, Pseudomonas aureofaciens 63–28 against the phytopathogen Rhizoctonia solani AG-4 on Petri plates and in soybean roots, growth response and induction of PR-proteins were estimated after inoculation
with P. aureofaciens 63–28 (P), with R. solani AG-4 (R), or with P. aureofaciens 63–28 + R. solani AG-4 (P + R). P. aureofaciens 63–28 showed strong antifungal activity against R. solani AG-4 pathogens in Petri plates. Treatment with P. aureofaciens 63–28 alone increased the emergence rate, shoot fresh weight, shoot dry weight and root fresh weight at 7 days after inoculation,
when compared to R. solani AG-4; P + R treatment showed similar effects. Peroxidase (POD) and β-1,3-glucanase activity of P. aureofaciens 63–28 treated roots increased by 41.1 and 49.9%, respectively, compared to control roots. POD was 26% greater in P + R treated
roots than R. solani treated roots. Two POD isozymes (59 and 27 kDa) were strongly induced in P + R treated roots. The apparent molecular weight
of chitinase from treated roots, as determined through SDS-PAGE separation and comparison with standards, was about 29 kDa.
Five β-1,3-glucanase isozymes (80, 70, 50, 46 and 19 kDa) were observed in all treatments. These results suggest that inoculation
of soybean plants with P. aureofaciens 63–28 elevates plant growth inhibition by R. solani AG-4 and activates PR-proteins, potentially through induction of systemic resistance mechanisms. 相似文献
11.
Dirk V. Charlson Sandeep Bhatnagar C. Andy King Jeffery D. Ray Clay H. Sneller Thomas E. CarterJr. Larry C. Purcell 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2009,119(4):587-594
As water demand for agriculture exceeds water availability, cropping systems need to become more efficient in water usage,
such as deployment of cultivars that sustain yield under drought conditions. Soybean cultivars differ in how quickly they
wilt during water-deficit stress, and this trait may lead to yield improvement during drought. The objective of this study
was to determine the genetic mechanism of canopy wilting in soybean using a mapping population of recombinant inbred lines
(RILs) derived from a cross between KS4895 and Jackson. Canopy wilting was rated in three environments using a rating scale
of 0 (no wilting) to 100 (severe wilting and plant death). Transgressive segregation was observed for the RIL population with
the parents expressing intermediate wilting scores. Using multiple-loci analysis, four quantitative trait loci (QTLs) on molecular
linkage groups (MLGs) A2, B2, D2, and F were detected (P ≤ 0.05), which collectively accounted for 47% of the phenotypic variation of genotypic means over all three environments.
An analysis of the data by state revealed that 44% of the observed phenotypic variation in the Arkansas environments could
be accounted for by these QTLs. Only the QTL on MLG F was detected at North Carolina where it accounted for 16% of the phenotypic
variation. These results demonstrate that the genetic mechanism controlling canopy wilting was polygenic and environmentally
sensitive and provide a foundation for future research to examine the importance of canopy wilting in drought tolerance of
soybean. 相似文献
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In this study we used the cuttings of Populus przewalskii Maximowicz as experimental material and sodium nitroprusside (SNP) as nitric oxide (NO) donor to determine the physiological
and biochemical responses to drought stress and the effect of NO on drought tolerance in woody plants. The results indicated
that drought stress not only significantly decreased biomass production, but also significantly increased hydrogen peroxide
content and caused oxidative stress to lipids and proteins assessed by the increase in malondialdehyde and total carbonyl
contents, respectively. The cuttings of P. przewalskii accumulated many amino acids for osmotic adjustment to lower water potential, and activated the antioxidant enzymes such
as superoxide dismutase, guaiacol peroxidase and ascorbate peroxidase to maintain the balance of generation and quenching
of reactive oxygen species. Moreover, exogenous SNP application significantly heightened the growth performance of P. przewalskii cuttings under drought treatment by promotion of proline accumulation and activation of antioxidant enzyme activities, while
under well-watered treatment the effect of SNP application was very little. 相似文献
15.
Siwaret Arikit Tadashi Yoshihashi Samart Wanchana Patcharin Tanya Ruangchai Juwattanasomran Peerasak Srinives Apichart Vanavichit 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,122(2):311-316
Vegetable soybean (Glycine max L.) is an important economic and nutritious crop in South and Southeast Asian countries and is increasingly grown in the
Western Hemisphere. Aromatic vegetable soybean is a special group of soybean varieties that produce young pods containing
a sweet aroma, which is produced mainly by the volatile compound 2-acetyl-1-pyrroline (2AP). Due to the aroma, the aromatic
vegetable soybean commands higher market prices and gains wider acceptance from unfamiliar consumers. We have previously reported
that the GmAMADH2 gene encodes an AMADH that regulates aroma (2AP) biosynthesis in soybeans (Arikit et al. 2010). A sequence variation involving a 2-bp deletion in exon 10 was found in this gene in all investigated aromatic varieties.
In this study, a codominant PCR-based marker for the aroma trait in soybeans was designed based on the 2-bp deletion in GmAMADH2. The marker was verified in five aromatic and five non-aromatic varieties as well as in F2 soybean population segregating for aroma. The aromatic genotype with the 2-bp deletion was completely associated with the
five aromatic soybean varieties as well as the aromatic progeny of the F2 population with seeds containing 2AP. Similarly, the non-aromatic genotype was associated with the five non-aromatic varieties
and non-aromatic progeny. The perfect co-segregation of the marker genotypes and aroma phenotypes confirmed that the marker
could be efficiently used for molecular breeding of soybeans for aroma. 相似文献
16.
The soybean aphid, Aphis glycines Matsumura is a new invasive pest of soybean in North America. We studied the ability of the existing predator community in soybean to reduce A. glycines establishment in field studies using either predator exclusion, open, or leaky cages that allowed aphid emigration but limited predation. Cages were infested with uniform initial densities of A. glycines adults and subsequent populations of aphids and predators were monitored over 24 h. The most abundant predators in these trials included the carabid beetles Elaphropus anceps (Le Conte), Clavina impressefrons Le Conte, Bembidion quadrimaculatum Say and spiders (Salticidae and Lycosidae). Foliar predators were less abundant and included; Harmonia axyridis Pallas, Coccinella septempunctata (L.), and Orius insidious (Say). Over the 2-year study, we found statistically significant predation on adult A. glycines in one out of six trials at 15 h and two out of six trials at 24 h. There was never significant evidence for predation of nymphs in any trial, however overall survival (adults + nymphs) was significantly reduced in one out of six trials at 15 h and three out of six trials at 24 h. Based on these results we suggest that generalist predators can be a significant but variable factor influencing the establishment of A. glycines populations in soybean. The impact of existing predator communities should be further investigated as a means of managing A.␣glycines populations in North American soybean production systems. 相似文献
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An efficient system of gene transformation is necessary for soybean [Glycine max (L.) Merrill] functional genomics and gene modification by using RNA interference (RNAi) technology. To establish such system, we improved the conditions of tissue culture and transformation for increasing the frequency of adventitious shoots and decreasing the browning and necrosis of hypocotyls. Adding N(6)-benzylaminopurine (BAP) and silver nitrate in culture medium enhanced the shoot formation on hypocotyls. BAP increased the frequency of the hypocotyls containing adventitious shoots, while silver nitrate increased the number of shoots on the hypocotyls. As a result, the number of adventitious shoots on hypocotyls cultured in medium containing both BAP and silver nitrate was 5-fold higher than the controls. Adding antioxidants in co-cultivation medium resulted in a significant decrease in occurrence of browning and necrosis of hypocotyls and increase in levels of beta-Glucuronidase (GUS) gene expression. Histochemical assays showed that the apical meristem of hypocotyls was the "target tissue" for Agrobacterium tumefaciens transformation of soybean. Gene silencing of functional gene by using RNAi technology was carried out under above conditions. A silencing construct containing an inverted-repeat fragment of the GmFAD2 gene was introduced into soybean by using the A. tumefaciens-mediated transformation. Several lines with high oleic acid were obtained, in which mean oleic acid content ranged from 71.5 to 81.9%. Our study demonstrates that this transgenic approach could be efficiently used to improve soybean quality and productivity through functional genomics. 相似文献
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Summary A characteristic phenotype of highly embryogenic explants along with the location of embryogenesis- and transformation-competent
cells/tissues on immature cotyledons of soybean [Glycine max (L.) Merrill.] under hygromycin selection was identified. This highly embryogenic immature cotyledon was characterized with
emergence of somatic embryos and incidence of browning/necrotic tissues along the margins and collapsed tissues in the mid-region
of an explant incubated upwards on the selection medium. The influences of various parameters on induction of somatic embryogenesis
on immature cotyledons following Agrobacterium tumefaciens-mediated transformation and selection were investigated. Using cotyledon explants derived from immature embryos of 5–8 mm
in length, a 1∶1 (v/v; bacterial cells to liquid D40 medium) concentration of bacterial suspension and 4-wk cocultivation
period significantly increased the frequency of transgenic somatic embryos. Whereas, increasing the infection period of explants
or subjecting explants to either wounding or acetosyringone treatments did not increase the frequency of transformation. An
optimal selection regime was identified when inoculated immature cotyledons were incubated on either 10 or 25 mgl−1 hygromycin for a 2-wk period, and then maintained on selection media containing 25 mgl−1 hygromycin in subsequent selection periods. However, somatic embryogenesis was completely inhibited when inoculated immature
cotyledons were incubated on a kanamycin selection medium. These findings clearly demonstrated that the tissue culture protocols
for transformation of soybean should be established under both Agrobacterium and selection conditions. 相似文献