世界栽培豌豆(Pisum sativum L.)资源群体结构与遗传多样性分析

【目的】评价国家种质库长期保存的国内外栽培豌豆(Pisum sativum L.)资源的遗传多样性水平,揭示其遗传多样性、等位基因和群体结构差异,据此评估其重要程度及价值,为中国豌豆资源研究策略和方向的正确选择、国内外资源的充分发掘利用和深入研究提供理论依据。【方法】利用21对豌豆多态性SSR引物,对来自国外五大洲66个国家和中国28个省(区、市)的1984份栽培豌豆进行SSR标记遗传多样性和群体结构分析;采用Structure2.2软件完成资源群体结构剖析、推断参试材料的合理组群数、确定每份参试材料的适当群体划入及其相关参数计算;利用NTSYSpc2.2d软件估算其遗传距离,进行主成分分析(PCA)并绘制三维空间聚类图;采用PopgeneV1.32估算种质群的等位位点分布等参数,利用FstatV2.9.3.2进行种质群间遗传多样性差异显著性测验。【结果】通过对国、内外栽培豌豆资源群间SSR等位基因数(NA)、有效等位基因数(NE)、有效等位基因所占比重(NE/NA)、等位基因丰度(AR)、基因多样性指数(GD)、Shannon's信息指数(I)的比较,发现除等位基因数(NA)外,国内资源的其它5个遗传多样性指标全面高于国外资源。在21个SSR基因位点中,国内、外资源群在7个位点间存在等位基因种类的差异。群体结构分析将1984份世界栽培豌豆资源划分成三大组群。组群A包含96.49%的国外栽培豌豆参试资源,可代表典型的国外栽培豌豆资源类型;组群B的资源88.18%来源于陕西和内蒙古,可代表中国典型的春播区栽培豌豆资源类型;组群C的资源52.05%源于中国秋播区,47.44%源于中国春播区,可代表中国秋播区和除陕西、内蒙古外的春播区栽培豌豆资源类型。组群间遗传多样性差异达到显著水平。PCA作图分析也明确显示世界栽培豌豆资源群体中存在3个边界明显的资源富集区(基因库I、II、III),且与三大组群的群体结构分析结果精确对应。【结论】国内资源的遗传多样性程度整体上超过国外资源,国外资源群体内个体间的差异程度平均高于国内资源。群体结构分析侦测到世界栽培豌豆资源中存在A、B、C共三大资源类群,类群间的遗传多样性差异达到了显著水平,且与PCA做图分析显示的3个边界明显的基因库间存在着精确对等关系:"类群A"几乎等同于"基因库I","类群B"几乎等同于"基因库II",而"类群C"几乎等同于"基因库III";基因库I由国外资源富集而成,基因库II由中国陕西和内蒙古资源富集而成,基因库III由中国秋播区资源和陕西、内蒙古以外的春播区资源富集而成,由此得出世界栽培豌豆由3个基因库构成的结论。国外栽培豌豆种质资源构成了"基因库I",国内栽培豌豆种质资源构成了"基因库II"和"基因库III",表明国内、外资源均很重要,但国内资源甚于国外。

Purifi cation, characterization and hypoglycemic activity of glycoproteins obtained from pea (Pisum sativum L.)

This study aimed to isolate and characterize the structures of glycoproteins from peas and determine their hypoglycemic activity.The crude pea glycoproteins(PGP)were extracted by hot water and purified by diethylaminoethyl(DEAE)-Sepharose chromatography and Sephadex G-100 size-exclusion chromatography in sequence.Then three main fractions were obtained,namely PGP1,PGP2 and PGP3,with molecular weights of 897615,846740 and 1194692 Da,respectively.The physical and chemical properties of the three fractions were evaluated and compared by Fourier transform infrared spectroscopy(FT-IR),nuclear magnetic resonance(NMR),scanning electron microscope(SEM),high performance liquid chromatography(HPLC)and other analytical techniques.The fraction PGP2 with the highest hypoglycemic activity,was screened using the Caco-2 monolayer cell model.It can inhibit the uptake of glucose in the small intestine,as well as the activities of maltase and sucrase.After simulated gastrointestinal digestion,PGP2 signifi cantly enhanced the inhibitory effect of α-glucosidase,and slightly reduced the inhibitory ability ofα-amylase.In summary,PGP2 possessed strong hypoglycemic activity after digestion.These results indicated that PGP2 has the potential to be developed into a functional food or natural medicine for the treatment of type 2 diabetes mellitus.

Large-scale evaluation of pea(Pisum sativum L.)germplasm for cold tolerance in the field during winter in Qingdao

As a cool-season crop, pea(Pisum sativum L.) can tolerate frost at the vegetative stage but experiences yield loss when freezing stress occurs at the reproductive stage. Cold-tolerance improvement of pea varieties is important for stable yield and expansion of the winter pea planting area. Under natural low-temperature conditions during winter in Qingdao,Shandong, China, we evaluated the cold tolerance of 3672 pea germplasm accessions in the field and categorized them as displaying high resistance(214), moderate resistance(835), or susceptibility(2623). The highly and moderately resistant genotypes were validated in the following year. We found that genotypes from the winter production region showed higher cold tolerance than genotypes from the spring production region. The accessions identified as having high levels of cold tolerance are recommended as potential genetic resources in cold-tolerance breeding of pea.

Rhizobium Inoculation and Micronutrient Addition Influence the Growth,Yield,Quality and Nutrient Uptake of Garden Peas(Pisum sativum L.)

Garden pea productivity and qualities are hampered in zinc(Zn),boron(B),and molybdenum(Mo)deficient soil.Thus,the combination of micronutrients(i.e.,Zn,B,and Mo)and rhizobium is necessary to increase the productivity and quality of garden peas,since this management for garden peas is neglected in Bangladesh.Therefore,the present study was made to assess the effectiveness of rhizobium inoculant singly or in combination with the micronutrients(i.e.,Zn,B,and Mo)on growth,yield,nutrient uptake,and quality of garden peas.Treatments were:T_(1)=Control,T_(2)=Rhizobium inoculation at 50 g/kg seed,T_(3)=T_(2)+Zn_(3)Mo1,T_(4)=T_(2)+B_(2)Mo1,T_(5)=T_(2)+Zn_(3)B_(2),T_(6)=T_(2)+Zn_(3)B_(2)Mo1 and T_(7)=Zn_(3)B_(2)Mo1.All treatments were arranged in a randomized complete block design and repeated all treatments in three times.The application of 3 kg Zn,2 kg B,and 1 kg Mo ha^(−1)with inoculation of Rhizobium at 50 g kg^(−1)seed(T_(6))facilitated to increase of 44.8%in the green pod and 29.7%seed yield over control.The same treatment contributed to attaining the maximum nodulation(25.3 plant^(−1)),Vitamin C(43.5 mg 100 g^(−1)),protein content(22.2%),and nutrient uptake as well as accumulation in garden peas.Among all treatment combinations,treatment T_(6)was found superior to others based on microbial activities,soil fertility,and profitability.The results of the study found that the application of 3 kg Zn,2 kg B,and 1 kg Mo ha^(−1)in combination with Rhizobium inoculation(50 g kg^(−1)seed)can improve the yield and quality of garden peas.The results of the study have the potential for the areas,where there is no use of Rhizobium inoculant or Zn,B,and Mo fertilizer for cultivation of garden pea.

镧对汞胁迫下豌豆(Pisum sativumL)幼苗生长及汞积累的影响

以培养 5d的豌豆 (PisumsativumL)幼苗为实验材料 ,研究了 10和 5 0mg·L- 1 的Hg胁迫下不同浓度La对豌豆幼苗生长的生理效应 ,测定并分析了La对Hg胁迫下豌豆幼苗根和叶细胞中Hg吸收及积累的影响。结果表明 ,合适浓度 (≤ 0 .0 10mg·L- 1 La)的La能在一定程度上缓解Hg胁迫所导致的叶绿体叶绿素和可溶性蛋白含量、光合速率和叶绿体Mg2 + ATPase、光合膜电子传递链活性及PSII活性的降低 ,减轻Hg胁迫对光合膜室温下吸收光谱和荧光光谱的影响 ;能有效增强或保护Hg胁迫下叶片细胞总抗氧化能力 ,降低细胞内Hg胁迫诱导超氧阴离子(·O2 - )产生的速率和膜脂过氧化产物 (MDA)的积累 ,延缓膜透性的增加。La对 10mg·L- 1 Hg胁迫的缓解能力较强 ,而对 5 0mg·L- 1 Hg的缓解效果较弱。最佳作用浓度为 0 .0 10mg·L- 1 。该浓度的La能有效的减少细胞对Hg的吸收和积累 ,特别是增加根细胞壁对Hg的束缚效应。超过一定浓度范围的La(≥ 0 .0 3 0mg·L- 1 )则加剧Hg的毒害作用。

马缨丹Lantana camara L.对豌豆Pisumsativum L.的化感作用

用不同体积分数的马缨丹Lantana camara L.水提取液在室内处理豌豆Pisum sativum L.种子和幼苗,以探讨前者对后者的化感作用.结果表明:马缨丹降低了豌豆种子活力,诱导根尖细胞产生较多的微核,破坏幼苗细胞膜的完整性.在其化感作用下,豌豆幼苗产生较多的丙二醛(MDA),同时提高了过氧化氢酶(CAT)和过氧化物酶(POD)的活性.显然,马缨丹对豌豆产生强烈的化感作用.

Heterogeneity in photosystem Ⅰfrom Pisum sativum L.

Studies have demonstrated that there are two types of PSI (PSI-1 and PSI-2) particles in some higher plants. These two types of PSI particles are proved to differ in organization and distribution on thylakoid membranes. PSI-1 mainly exists in the non-partition regions of thylakoids, while PSI-2 mainly in the partition regions.

Rhizogenesis of Two Species Fabaceae: <i>Cicer arietinum</i>L. and <i>Pisum sativum</i>L.

Our work of comparative study on the biomorphological, rhizogenic level of Chickpea (Cicer arietinum) and Pea (Pisum sativum), includes successively the following results: The in-vitro culture done, showed strains, which can reach 6.5 mm in the medium Nutrient agar (NA) for Cicer arietinum 6 mm of strain were recorded in the PDA culture medium for Pisum sativum. The best strain is obtained in temperature of 20&#8451;(ambient). According to the results of the rhizogenesis, we notice that the nutrient agar (NA) medium responds better that the PDA medium for Cicer arietinum. Meanwhile, the medium PDA brings a better reaction (response) compared with the nutrient agar medium concerning Pisum sativum.

Physiological Responses of Pea Plants to Salinity and Gibberellic Acid

Pea is a seed legume.It is rich in cellulose fibre and protein.It is also a significant source of minerals and vitamins.In this paper,we set out to better characterize the physiological responses of Pisum sativum L.to the combined effects of NaCl,100 mM and gibberellins(GA3).Our analysis revealed that NaCl caused a decrease in growth resulting in a reduction in root elongation,distribution and density,leaf number and leaf area,and a decrease in dry matter of roots and shoots.However,the contribution of GA3 in the salty environment induced an increase in these different parameters suggesting an improving effect of this hormone on growth of pea in presence of salt.NaCl also led to a disturbance of the photosynthetic machinery.Indeed,level of chlorophyll pigments(a and total)and photosynthetic activity were decreased compared to the control plants.However,the exogenous supply of GA3 restored this decrease in net CO_(2) assimilation,but not in chlorophyll content.Additional analyses were performed on the effect of salinity/GA3 interaction on osmolytes(soluble sugars and starch).Our results showed an increase in sugars and a decrease in starch in the presence of 100 mM NaCl.The salt-GA3 combination resulted in compensation of soluble sugar contents but not of starch contents,suggesting a beneficial effect of GA3 under saline stress conditions.Level of three main polyamines putrescine,spermidine,and spermine increased significantly in all organs of salt-treated plants.

外源ALA对盐胁迫下豌豆幼苗生理特性的影响

在温室条件下研究外源ALA对盐胁迫下豌豆幼苗生理特性的影响.在100mmol/L NaCl盐胁迫下,研究0.001～1mmol/L ALA对豌豆幼苗生理特性的影响.100mmol/L NaCl盐胁迫导致豌豆幼苗氧化损伤,SOD等保护酶活性升高;0.1 mmol/L ALA处理对盐胁迫下的豌豆幼苗有显著的保护作用,使叶片MDA含量降低25.87%,SOD,POD和CAT活性分别提高28.74%,20.68%和36.83%,幼苗株高、鲜物质量分别提高0.975,0.15倍,从而增强幼苗的抗盐性.0.1mmol/L ALA能缓解盐胁迫下豌豆幼苗的氧化损伤,显著提高SOD等保护酶活性.

Response of Endogenous Salicylic Acid and Jasmonates to Mechanical Wounding in Pea Leaves

The roles of on endogenous jasmonates （JAs） and salicylic acid （SA） in wounding response were investigated. Pea （Pisum sativum L.） seedlings were treated with three different methods including mechanical wounding, JAs application, and SA application. The contents of endogenous JAs and SA, as well as the activities of the related enzymes were detected by enzyme-linked immunosorbent assay （ELISA）, high performance liquid chromatography （HPLC）, and spectrophotometer, respectively. The results showed that endogenous JA rapidly accumulated within 30 min after wounding. The increase in the activities of both lipoxygenase （LOX） and allene oxide synthase （AOS） lagged behind JAs burst. A second slight increase in JAs level was observed at 24 h after wounding treatment, and at the same time point, higher activities of LOX and AOS were also detected. Endogenous free SA content decreased accompanied with JAs burst. Effects of exogenous JA application were similar to those of wounding treatment on endogenous SA level and phenylalanine ammonia lyase （PAL） activity, whereas exogenous SA application led to the significant inhibition of LOX and AOS activities and the decrease of endogenous JAs level at the early stage of treatment. It is thus suggested that JAs burst and SA decrease in early response to wounding may constitute an important mechanism by which plant starts the related defense reaction and adapts to wounding stress.

Effects of Wounding and Exogenous Jasmonic Acid on the Peroxidation of Membrane Lipid in Pea Seedlings Leaves

The changes of malondialdehyde （MDA）, H2O2, and O2^7 content, or the activities of superoxide dismutase （SOD）, catalase （CAT）, ascrobate peroxidase （APX）, peroxidase （POD）, phenylalanine ammonia lyase （PAL）, and polyphenol oxidase （PPO） in pea seedlings （Pisum sativum L.） under wounding and treatment of exogenous jasmonic acid （JA） were investigated. The results showed that the activities of both phenylalanine ammonia lyase （PAL） and polyphenol oxidase （PPO） were significantly increased by wounding and application of JA. The metabolism of reaction oxidative species （ROS） was enhanced, especially O2^7 and H2O2 appeared to rapidly increase. The activities of antioxidant enzymes such as SOD, CAT, APX and POD were also increased. Treatment of JA of 1 or 10 μmol L^-1 could effectively induce plant defense response, and thus decrease the peroxidation of cell membrane lipid. However, high concentration of JA （100 μmol L^-1） resulted in unbalance of metabolism of ROS and promoted the peroxidation of cell membrane lipid. We thus suggested that JA, under the suitable concentration, could induce defense response of pea seedlings to wounding.

Comparative Genomics for Gene Isolation in Legumes

The similarity in gene order between closely related taxa suggests that genomic information from model systems should facilitate gene isolation and characterization in target crops. If this is the case, a great deal of

富铬(Ⅲ)豌豆制备及其含铬蛋白组分生物活性研究

目的 制备富铬(Ⅲ)豌豆(Pisum sativum L.),并探究富铬豌豆蛋白的体外生物活性,为新型有机铬产品开发提供参考。方法 以豌豆为实验材料,通过电感耦合等离子体质谱法测定不同形态铬的含量,以有机铬含量为指标评价各因素水平对铬(Ⅲ)的生物富集效应影响;对比不同萌发生长周期的富铬豌豆铬含量及蛋白质铬含量。并通过体外降糖、降脂及抗氧化实验评价富铬豌豆蛋白的生物活性。结果 豌豆种子浸泡的最佳条件为:铬离子浓度为1.32 mmol/L的三氯化铬溶液在20℃浸泡12 h,该条件下,富铬豌豆铬含量约为未富铬豌豆铬含量的930倍,富铬豌豆中有机铬的含量为(170.65±2.61)μg/g,有机化程度达87%;萌发过程中富铬豌豆的铬含量呈下降趋势,但其蛋白质铬含量在萌发第4 d最高,为(82.15±0.45)μg/g,随后下降。生物活性研究表明富铬豌豆蛋白具有良好的体外降糖、降脂及抗氧化活性。结论 利用豌豆通过生物富集方法将无机铬转化为有机铬是切实可行的,富铬豌豆蛋白具有优于普通豌豆蛋白的体外降糖、降脂及抗氧化活性。

纳米硒微肥的形貌分析及对豌豆幼苗生长的影响

为制备筛选易于吸收的纳米硒微肥,利用化学还原法制备纳米硒,通过不同干燥处理和扫描电镜技术研究纳米硒的形貌特征、尺寸及稳定性。将纳米硒与豌豆种子拌种后进行盆栽试验,测定豌豆幼苗的硒吸收量和硒转化率。结果表明,透射电镜和扫描电镜下,冷冻干燥制备的纳米硒为分散均匀、尺寸均一的球状颗粒,平均粒径约为28 nm。当拌种剂量(纳米硒/豌豆种子)在0.25~2.50 mg·g^(-1)范围内,纳米硒微肥对豌豆幼苗生理指标的影响较小。豌豆幼苗根系硒含量均达到极显著水平,地上部分硒含量在2.00和2.50 mg·g^(-1)拌种剂量时显著高于对照和其他处理;在1.00~2.50 mg·g^(-1)范围内,硒吸收量存在显著差异;在0.25~2.50 mg·g^(-1)范围内,硒转化率呈先降低后增加的趋势。综上,冷冻干燥制备法可获得分散均匀、尺寸均一的纳米硒微肥。拌种剂量控制在0.25~2.50 mg·g^(-1)范围内时,豌豆幼苗地下部分对纳米硒的吸收率大于硒转化率,可采用冷冻干燥法制备纳米硒微肥用于豌豆生产。

豌豆营养价值与开发利用研究进展

对菜用豌豆(Pisum sativum L.)和干子粒豌豆中营养成分的研究进行了综述,详细归纳了加工因素对菜豌豆营养活性物质变化的影响,并总结了豌豆作为饲料在养殖产业中的利用价值,以及近年来其药理学特殊活性成分在人类疾病治疗中的开发应用。结合当下的研究热点和前景对开拓特色功能豌豆的研究进行了探讨和展望。

中国豌豆地方品种SSR标记遗传多样性分析

利用21对豌豆多态性SSR引物,对来自全国春、秋播区19省区市的1221份豌豆地方品种进行遗传多样性分析,共扩增出104条多态性带,每对引物平均扩增出4.95个等位变异,其中有效等位变异占62.52%。省份间SSR等位变异分布均匀,但是省份间有效等位变异数、Shannon’s信息指数(Ⅰ)差异明显,省籍资源群间遗传多样性差异显著。遗传多样性以内蒙古资源群最高,甘肃、四川、云南和西藏等资源群其次,辽宁资源群最低。PCA三维空间聚类图揭示,我国豌豆地方品种资源分化成3个基因库,基因库Ⅰ主要由春播区的内蒙古、陕西资源构成,基因库Ⅱ主要由秋播区最北端的河南资源构成,基因库Ⅲ主要由除上述省份之外的其他省区市的资源构成。UPGMA聚类分析表明,不同省份资源群间的遗传距离变化范围为5.159～27.586,中国豌豆地方资源据此聚类成2个组群8个亚组群,与3个基因库的聚类结果相呼应。聚类结果显示,我国豌豆地方品种资源群间遗传距离与其来源地生态环境相关联。

国外栽培豌豆遗传多样性分析及核心种质构建

从111对备选SSR引物中筛选出能扩增出清晰稳定单一带的多态性引物21对及其最佳退火温度,并优化了豌豆SSR标记实验体系。利用上述引物,对来自于67个国家的731份豌豆栽培种质(Pisum sativum L.)进行遗传多样性分析与核心种质构建。共扩增出109条多态性带,每对引物平均扩增出5.19个等位变异。SSR等位变异在各大洲间分布不均匀,有效等位变异数、Shannon's信息指数(I)洲际间差异明显。各大洲资源群间遗传多样性差异显著,其中亚洲最高(I=1.1753),欧洲其次(I=1.1387),俄罗斯联邦(I=1.0285)、美洲(I=1.0196)、非洲(I=0.9254)、大洋洲(I=0.8608)依次降低。利用Popgene1.32软件,依豌豆栽培资源洲际间Nei78遗传距离可聚类成2个组群和4个亚组群;基于Structure 2.2软件分析,国外栽培豌豆资源实际由3大类群组成,并与Popgene 1.32聚类结果呼应得较好。上述两种分析方法均表明,国外栽培豌豆类群的遗传多样性与其地理分布相关。设计并实践了一套基于Structure分析的科学可靠、逻辑性强的核心种质构建标准化方案,并依此构建了一套以6.57%的资源(48份)涵盖总体84.4%等位变异的国外栽培豌豆核心种质。

271份豌豆种质资源农艺性状遗传多样性分析

以来自五大洲57个国家和地区的271份豌豆资源为材料,利用主成分分析和聚类分析的方法,评价其2个质量性状和7个数量性状的遗传变异水平。结果表明,参试资源的农艺性状具有丰富的遗传多样性。其中遗传多样性指数最高的是始荚节位(2.0590),其次是主茎节数(2.0421);性状变异系数最大的是小区产量(64.874%),其次是百粒重(61.870%)。采用SPSS 22.0软件对参试资源7个数量性状的主成分分析结果表明,前3个主成分因子累计贡献率达66.021%,Ward法聚类将参试的271份豌豆资源划分为4大类群,其中第Ⅱ类群属于高秆、大粒、高产种质,具有很高的丰产潜力,为我国杂交育种亲本选择提供了原材料。对取材数大于等于5的来自于四大洲17个国家的参试资源进一步分析发现,不同地理区域资源间遗传变异显著,其中印度的遗传多样性指数最高(1.7533),巴基斯坦的最低(0.9685),波兰的变异系数和遗传多样性都较高。综合分析国外种质资源的农艺性状,有助于我国豌豆育种项目亲本选配。

机械伤害和茉莉酸对豌豆幼苗膜脂过氧化的影响

切割后离体芽苗极易受病菌侵染而发生腐烂变质,严重影响着当前芽菜采后运输和销售。为此,本试验以豌豆芽苗为试材,研究了机械伤害和外施不同浓度(±)茉莉酸(JA)处理对豌豆幼苗膜脂过氧化的影响。结果表明,伤害和1～100ìmol·L-1JA处理可以显著提高苯丙氨酸解氨酶(PAL)和多酚氧化酶(PPO)活性,同时使活性氧代谢加强,O2·产生速率和H2O2含量迅速增加,相应的抗氧化酶(SOD、CAT、APX、POD)活性提高。-适宜浓度JA(1～10ìmol·L-1)处理可有效诱导防御反应产生,同时,通过提高抗氧化酶活性使膜脂过氧化程度减轻;高浓度JA(100ìmol·L-1)处理造成植物活性氧代谢失调,膜脂过氧化加剧。综合分析认为,10ìmol·L-1JA是诱导豌豆幼苗防御反应产生的适宜处理浓度。