普通菜豆(Phaseolus vulgaris L.)AOX基因家族鉴定与表达分析

交替氧化酶(Alternative oxidase,AOX)是一种末端氧化酶,在维持植物正常生长代谢速率和代谢平衡以及抵御逆境胁迫中发挥了一定作用。AOX家族成员已经在很多植物物种中被鉴定,但是在普通菜豆(Phaseolus vulgaris L.)中却未见报道。研究对普通菜豆中AOX家族成员进行了鉴定和分析,并进一步解析了AOX基因家族成员的功能及调控机制。结果表明,基于普通菜豆参考基因组中共挖掘得到了4个AOX家族成员,通过进化关系、保守基序(Motifs)、基因结构分析,将各成员划分成4个亚族。此外,顺式作用元件、共线性与盐胁迫下的表达量分析结果表明,AOX成员可能在植物的非生物胁迫响应中发挥了作用。为后续深入研究普通菜豆中AOX成员的功能奠定了理论基础。

A novel pathogen Fusarium cuneirostrum causing common bean(Phaseolus vulgaris)root rot in China

Several fungal pathogens cause root rot of common bean,among which Fusarium spp.are the most common pathogens causing Fusarium root rot(FRR)worldwide.FRR has been becoming an increasingly severe disease of common bean in China,but the species of Fusarium spp.have remained unclear.Thus,this study was performed to identify the pathogen causing common bean root rot in Liangcheng County,Inner Mongolia,China.Nineteen Fusarium-like isolates were obtained after pathogen isolation and purification.The pathogenicity test indicated that eight isolates caused severe disease symptoms on common bean,while 11 other isolates were not pathogenic.The eight pathogenic isolates,FCL1–FCL8,were identified as Fusarium cuneirostrum by morphological characterization and phylogenetic analysis using partial sequences of EF-1α,ITS,28S,and IGS regions.Host range test showed that the representative F.cuneirostrum isolate FCL3 was also pathogenic to mung bean,while not pathogenic to adzuki bean,chickpea,cowpea,faba bean,pea,and soybean.Moreover,50 common bean and 50 mung bean cultivars were screened for resistance to FRR,and seven highly resistant or resistant cultivars of common bean were identified,while no resistant cultivars of mung bean were screened.This study revealed that F.cuneirostrum was one of common bean FRR pathogens in Inner Mongolia and it could induce mung bean root rot as well.To our knowledge,this is the first report of F.cuneirostrum causing FRR of common bean in China.

Transcriptome Analysis of Molecular Mechanisms Underlying Phenotypic Variation in Phaseolus vulgaris Mutant‘nts’

The phenotype of a common bean plant is often closely related to its yield,and the yield of plants with reduced height or poor stem development during growth is low.Mutants serve as an essential gene resource for common bean breeding genetic research.Although model plants and crops are studied to comprehend the molecular mechanisms and genetic basis of plant phenotypes,the molecular mechanism of phenotypic variation in common beans remains underexplored.We here used the mutant‘nts’as material for transcriptome sequencing analysis.This mutant was obtained through 60Co-γirradiation from the common bean variety‘A18’.Differentially expressed genes were mainly enriched in GO functional entries such as cell wall organization,auxin response and transcription factor activity.Metabolic pathways significantly enriched in KEGG analysis included plant hormone signal transduction pathways,phenylpropanoid biosynthesis pathways,and fructose and mannose metabolic pathways.AUX1(Phvul.001G241500),the gene responsible for auxin transport,may be the key gene for auxin content inhibition.In the plant hormone signal transduction pathway,AUX1 expression was downregulated and auxin transport across the membrane was blocked,resulting in stunted growth of the mutant‘nts’.The results provide important clues for revealing the molecular mechanism of‘nts’phenotype regulation in bean mutants and offer basic materials for breeding beneficial phenotypes of bean varieties.

Nutritional and Mineral Variability in 52 Accessions of Common Bean Varieties(Phaseolus vulgaris L.)from Madeira Island

The Phaseolus vulgaris L. is one of the traditional and most important leguminous crops in the Island of Madeira. The island’s bioclimatic tiers, agro-ecological environment and traditional farming practices had a great influence on the evolution of regional bean landraces. The variability of the nutritional and mineral seeds composition of 59 accessions of the Madeiran landraces, standard and commercial varieties was evaluated. Wide ranges of variability in biochemical parameters were reported among the Madeiran landraces, being the best sources of protein and mineral nutrition, according to the statistical results and literature comparative evaluation. Specifically, the content (g per 100 g DW) of ash ranged from 3.64-5.67, lipids from 0.57-2.86, protein from 18.55-29.69, starch from 23.40-52.65, soluble sugars from 2.97-6.84, while content of dry matter was from 83.35-93.55. The seeds also contained (per 100 g DW) between 2.55-4.83 g N, 0.30-7.50 g P, 1.30-2.49 g K, 0.10-0.18 g Mg, 4.10-10.00 mg Fe, 50.0-1.40 mg Cu, 2.20-5.00 mg Zn, 0.90-3.80 μg Mn and 0.20-2.40 μg B. This variability implies that the screened germplasm could serve as a source for breeding new varieties with improved biochemical and nutritional traits or could be highly recommended to meet specific dietary requirements. The cultivar Vaginha Grossa (ISOP 713) revealed low carbohydrate content that could be a good food choice for diabetics, while cultivar Vermelho (ISOP 724) bean should be offered as a valuable alternative source of protein and minerals in the local diet.

Rapid,Efficient and High-Performance Protocol for Agrobacterium rhizogenes-Mediated Hairy Root Transformation of the Common Bean Phaseolus vulgaris

A rapid, efficient and high-performance transformation protocol employing Agrobacterium rhizogenes was developed for the common bean Phaseolus vulgaris. In this study, we examined competencies of various protocols to induce and explants that respond to hairy root transformation in bean plants. Utilizing young seedlings with severed radicles/hypocotyls, we developed a highly efficient procedure for achieving hairy root transformation frequencies as high as 100% as visualized by GUS reporter gene expression system. Transgenic hairy roots in these young composite plants were susceptible to nodulation by rhizobia, and form an excellent system for high throughput genomic analysis to study root biology and endosymbiosis in common bean.

Differential Effect of Aluminium on Enzymes of Nitrogen Assimilation in Excised Bean Leaf Segments

Aluminium is a potent toxicant in acidic soils. The present study was taken up to analyze the effects of Al on enzymes of nitrogen assimilation in excised bean (Phaseolus vulgaris) leaf segments so as to gain an insight of the mechanism involved. Supply of 0.001 to 0.1 mM AlCl3 to excised bean leaf segments affected the in vivo nitrate reductase activity differently in the presence of various inorganic nitrogenous compounds, being inhibited with 5 mM ammonium nitrate and 10 mM ammonium chloride but enhanced with 10 mM potassium nitrate. Al effect with 50 mM KNO3 varied with time, showing an increased activity at shorter duration, but decreased at longer duration. Al effect on in vivo NRA was dependent upon the nitrate concentration, thus, inhibiting it at 0, 1 and 50 mM KNO3, while increasing at 2 and 10 mM. Further, saturating and non-saturating effects were observed in the absence and presence of Al. Al supply influenced the in vitro NRA also, being increased at 10 mM, but decreased at 50 mM KNO3. Supply of Al to excised leaf segments substantially inhibited the glutamate dehydrogenase activity in the absence as well as presence of 5 mM NH4NO3 but increased the glutamate synthase activity. Inhibition of specific glutamate dehydrogenase activity by Al supply was also observed. However, specific glutamate synthase activity was increased in the presence of NH4NO3 only. The experiments demonstrated that effect of supply of aluminium on in vivo nitrate reductase activity depended upon nitrogenous source as well as nitrate concentration and it exerted reciprocal regulation of glutamate dehydrogenase and glutamate synthase activities, which depended upon N supply too.

Can Improve Iron Biofortification Antioxidant Response,Yield and Nutritional Quality in Green Bean?

The aim of this study was to evaluate the effect of iron biofortification on antioxidant response, yield and nutritional qualityof green bean (Phaseolus vulgaris L.) under greenhouse conditions. Fe was applied using two forms (FeSO4 and Fe-EDDHA) at four doses of application (0, 25, 50 and 100 μm) added under a hydroponic system, and were tested over a period of 40 days. The Fe content was assessed in seeds, as well as the activity of antioxidant enzymes, production of H2O2, yield and nutritional quality. The results being obtained indicated that the accumulation of Fe in bean seeds enhanced with the application of Fe-EDDHA at the dose of 25 μm. This demonstrated that low Fe application dose was enough to increase Fe levels in seeds of common bean. In addition, Fe-EDDHA application form at 50 μmol was the best treatment to improve crop yield. Respect to antioxidant system, chelated form of Fe (Fe-EDDHA) was more effective in the activation of antioxidant enzymes (CAT, SOD and GSH-PX), and a lower content of H2O2 in green bean seeds. Finally, to raise the Fe concentration in bean under biofortification program was a promising strategy in cropping systems in order to increase the ingestion of iron and antioxidant capacity in the general population and provided the benefits that this element offered in human health.

Meiotic behavior and pollinic viability in bean cultivars

The aims of this study were to determine the meiotic behavior and to estimate pollen grains via-bility in bean(Phaseolus vulgaris L.)cultivars.Flower buds were collected during different developmental stages of the Mesoamerican bean cultivars IAPAR 44,Guapo Brilhante,BRS Expedito,BRS Valente,Guateian 6662 and Pérola,and the Andean bean cultivar Iraí,grown in a greenhouse.The meiotic index was determined by anther squashing of material fixed in absolute ethanol-glacial acetic acid(3:1)and stained with acetic orcein.No meiotic abnormalities were observed and the meiotic indices were high for all cultivars,indicating that the mismatch generated during crosses is not related to any meiotic changes.Estimation of pollen viability was made by comparing acetic orcein staining vs.Alexander’s reactive:pollen viability was high in all cultivars with either stain,but was significantly higher when using the acetic orcein stain(>99%).Though some cultivar showed a significantly smaller pollen size,the range of variation among cultivars was low(means’range 51-66μm).

Extraction and Solubilization Methods Suitable for Proteomic Analysis of Common Bean Seed of Ivory Coast

The extraction and solubilization of proteins from seeds of Phaseolus vulgarisms for two-dimension polyacrylamide gel electrophoresis (2D-PAGE) and analysis by mass spectrometry are very sensitive procedures. In this study, we used two methods of extraction and solubilization of proteins, the urea/thiourea method and the trichloroacetic acid (TCA)/acetone precipitation method, in order to determine their effectiveness in separating proteins from bean seeds by 2D-PAGE. In both methods, proteins were well separated by 2D PAGE with minor variations in the protein pattern. These two extraction methods showed that it was possible to separate hundreds of very resolvent proteins by 2D electrophoresis. A protein spot was selected on the 2D-PAGE gel, digested with trypsin and analyzed by mass spectrometry (LCMS/ MS). The results suggest that thiourea/urea and TCA methods were effective and reliable for the extraction and solubilization for 2D analysis of proteins from seeds of Phaseolus vulgaris .

Weed Control with Sulfentrazone plus a Low Rate of Imazethapyr in White Bean

Ontario dry bean (Phaseolus vulgaris L.) growers have few options for broadleaf weed control. Sulfentrazone is a group 14 herbicide that provides good control of several common Ontario weed species, and would provide another mode of action for broadleaf weed control if registered for use in Ontario dry beans. Five field studies were conducted during 2014 and 2015 to determine if a low dose of imazethapyr added to a tank mix of sulfentrazone + s-metolachlor would improve broadleaf control in white bean. Sulfentrazone (140 and 210 g·ai·ha-1) was mixed with imazethapyr and s-metolachlor and evaluated at 2 and 4 weeks after crop emergence for crop injury. Weed control was assessed visually at 4 and 8 weeks after herbicide application (WAA), and weed stand counts and biomass were determined at 8 WAA. Seed moisture and yield were determined at harvest. At 8 WAA, sulfentrazone (140 g·ai·ha-1) controlled pigweed species, common ragweed, common lambsquarters, wild mustard, barnyard grass and green foxtail 100%, 4%, 100%, 2%, 86% and 62%, respectively. The addition of imazethapyr (37.5 g·ai·ha-1) to sulfentrazone (140 g·ai·ha-1) improved the control of common ragweed, wild mustard and green foxtail by 19%, 98% and 33%, respectively. The three-way tank mix of sulfentrazone (140 g·ai·ha-1) plus s-metolachlor (1050 g·ai·ha-1) plus imazethapyr (37.5 g·ai·ha-1) controlled pigweed species, common ragweed, common lambsquarters, wild mustard, barnyard grass and green foxtail 100%, 35%, 100%, 100%, 96% and 100%, respectively. The tank mixes evaluated caused unacceptably high levels of crop injury;this study does not support the registration of sulfentrazone plus s-metolachlor + imazethapyr for use in Ontario white bean.

Decomposition of the interaction of common black bean group genotypes with the environment

The purpose of this paper was to assess which environmental factor(s) (location, sowing season or years) in the common black bean genotypes interaction was more expressive and to verify the existence of genotypes with broad and/or specific adaptations to sowing season. Data of grain yield of thirteen genotypes were tested in twelve trials of value of cultivation and use during 2003 and 2004 in the south-central region of Brazil. Analyses of variance were performed with the decomposition of genotypes × environments interaction, which allowed a partial isolation of factors, and subsequently, eight trials were used which allowed a complete isolation of factors. The most important interactions were genotypes × years (R2 = 2.5%) and genotypes × locations (R2 = 1.9%), while genotypes × sowing seasons was the least important (R2 = 1.4%). Thus, it is more important to evaluate the genotypes in different locations and years than at different seasons. Most genotypes showed wide adaptation, but some lines showed strong specific adaptation.

Effect of Osmotic Stress on the Initial Development of Bean Seedlings

One of the methods to determine the tolerance of plants to water stress is the observation of germination of seeds under stress induced in laboratory conditions. Thus, the aim was to evaluate the response of four common bean cultivars to water deficit simulated under laboratory conditions. Two common landrace genotypes acquired from farmers (FORT 07 and FORT 08) and two commercial cultivars, IAPAR 81 and Serrano were evaluated. The different levels of water deficit were induced with mannitol solution. The experimental design was completely randomized and a factorial scheme 5 × 4 (osmotic potential x genotypes), with four replications. The characteristics: primary root protrusion, seed vigor, normal and abnormal seedling, seedling and root length, fresh and dry weight of seedling and root were evaluated. By reducing the osmotic potential seedling vigor bean was more affected than the primary root protrusion. Regardless of cultivar, there was a significant and progressive reduction in the normal seedlings percentage from potential -0.3 MPa. The IAPAR 81 and FORT 07 cultivars showed higher tolerance to water deficit compared to other cultivars.

内蒙古普通菜豆枯萎病病原菌鉴定

普通菜豆(Common bean),俗称芸豆(Kidney bean),为豆科(Fabaceae)菜豆属一年生草本植物,营养丰富,蛋白质含量高,既是蔬菜又是粮食,是重要的农副产品。内蒙古自治区是我国种植普通菜豆的主要地区之一。随着普通菜豆栽培面积以及连作年限不断增加,病虫害问题日益突出,尤其是枯萎病,严重影响了普通菜豆的产量和品质,已成为限制普通菜豆产业可持续发展的主要因素之一。

贵州普通菜豆抗锈病及抗白粉病基因SCAR标记鉴定

利用来自于普通菜豆11个SCAR标记对54份来源于贵州省11个县的普通菜豆资源的Ur-3、Ur-4、Ur-5、Ur-6、Ur-7、Ur-11、Ur-13和Ouro Negro等8个抗锈病基因以及SAU5、SS18、SF6Em3等3个抗白粉病基因进行了分子标记鉴定。结果表明:参试材料一般携带其中1~11个抗性基因,品种间差异较大;各基因频率存在较大差异(频率0~100%),抗锈病基因Ur-5(88.59%)、Ur-7(72.22%)、Ur-11(75.93%)和抗白粉病基因SF6Em3(64.81%)、S18(100.00%)在参试材料中携带频率高,为优势基因。Ur-5、Ur-7、Ur-11和S18的在贵州不同地区分布较为均衡,而其余基因具有较为明显的区域特性。在54份材料中,11个测试基因标记组合形成了46种基因型,多样性较丰富。本研究明确了54份参试菜豆资源所含的抗锈病及抗白粉病基因型,分析了不同抗病基因在贵州各地的分布规律,筛选出了抗锈病及抗白粉病基因聚合体的菜豆资源,为贵州菜豆资源在育种中的应用奠定了基础。

菜豆DUS测试标准品种SSR遗传多样性分析

为了对《植物新品种特异性,一致性和稳定性测试指南菜豆》标准品种选用的合理性进行评价,为测试指南进一步的修订和完善提供分子层面的参考依据,本研究对菜豆DUS测试标准品种的遗传多样性进行分析。结果显示,33对核心引物在26个标准品种中共检测到214个等位变异,平均每对引物检测到了6.49个等位变异,该套引物的Shannon's信息指数在0.5158~2.1257之间,均值是1.2957;多态信息含量(PIC)处于0.2480~0.8609之间,平均值为0.6040。26个标准品种的遗传相似系数在0.03~0.85之间,平均值为0.31,表明品种间遗传多样性较丰富,标准品种选择较为合理。其中仅‘双阳大花皮’和‘大家花雀蛋间’的遗传相似系数大于0.8,且二者在测试指南中均只指示一个性状,可以根据这一结果开展进一步田间对比试验,尝试减少标准品种数量,提高DUS测试效率。

基于SSR及ISSR标记的菜豆遗传多样性分析

本研究以70份菜豆材料为研究对象,利用7对SSR和13对ISSR引物,对其进行遗传多样性分析,以期为菜豆的分子标记辅助育种研究提供理论依据。结果表明:SSR-PCR共扩增出52条带,其中多态性带39条,多态性比率为75%;ISSR-PCR共扩增出117条带,其中多态性带96条,多态性比率为82.1%。UPGMA法聚类均可将试验材料分为4个类群,7个亚类,SSR+ISSR分析得到菜豆材料遗传相似系数在0.222~0.944之间,平均值0.625,反映出试验材料具有较高的遗传多样性。采用两种分子标记相结合的方法研究70份菜豆种质资源间的亲缘关系,获得可靠的聚类结果,材料具有丰富的种质遗传多样性,结果可对保护菜豆种质资源和研究利用提供参考。

The Diversity of Four Anti—nutritional Factors in Common Bean

Anti-nutritional factors such as lectins, saponin, trypsin inhibitor and phytic acid are endogenous substances in the common bean(Phaseolus vulgaris L.). In this study, the contents or activities of these anti-nutritional factors in fresh pods were detected in 56 selected cultivars. The results revealed significant difference within each factor in the tested cultivar population. The mean value of lectin content and the activity of trypsin inhibitor were 1.743 mg · g^(-1)and 1.680 mg · g^(-1)respectively. Their coefficients of variation(CV) were both more than 100% and each of the cultivar frequency distribution curve showed a main peak, but the discontinuous distributions in the extremely high and low areas indicate hierarchic cultivars. However, the mean contents of saponin and phytic acid were 3.730 mg · g^(-1)and 3.102 mg · g^(-1), respectively, with CV less than 41%.Each showed a main peak in its normal distribution curve and low frequency continuous distribution in dual tails. Meanwhile, statistic analysis demonstrated a positive correlation between the lectin content and trypsin inhibitor activity in fresh pods. Furthermore, all 56 tested cultivars were clustered into three groups based on their four anti-nutritional factor levels: 80% of them into medium level group, and 12% of them into low level group. The endogenous edible toxic compounds, such as lectin and trypsin inhibitor, are closely related to insect resistance in the field.This study suggests that it is possible to screen the cultivars containing less lectin and other factors but with reduced pest resistance in the field.

Synthesis of an eco-friendly nanocomposite fertilizer for common bean based on carbon nanoparticles from agricultural waste biochar

The burning of agricultural waste is a major cause of environmental pollution.In this study,we sought to prepare biochar from agricultural waste as a source material for the preparation of carbon nanoparticles(CNPs).Surface morphology,hydrodynamic particle size,and purity and crystallinity of CNPs were extensively investigated using transmission electron microscopy(TEM),zeta sizing,and X-ray diffraction(XRD)spectroscopy,respectively.The CNPs were subsequently immersed in a solution of potassium nitrate(KNO_(3))to prepare a CNPs/NK nanocomposite(CNPs loaded with nitrogen(N)and potassium(K))as a nanocomposite fertilizer for common bean(Phaseolus vulgaris L.).The CNPs/NK nanocomposite was sprayed as a foliar fertilizer at 0,10,20,30,and 40 mg L^(-1) on common bean plants 25 d after sowing on a farm in Shebin El-Kom,El-Monifia,Egypt.The growth,yield,and quality of common bean were investigated during two successive growing seasons(2017 and 2018).The highest seed yields of 2.04 and 2.01 t ha^(-1) and the highest values of growth parameters including plant height of 61.5 and 59.2 cm,number of leaves per plant of 35 and 35,number of flowers per plant of 83.3 and 82.7,and plant fresh weight of 148.7 and 152.8 g plant^(-1) were obtained when using the CNPs/NK nanocomposite at a concentration of 20 mg L^(-1) during the 2017 and 2018 growing seasons,respectively.