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.

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.

150份豌豆(Pisum sativum L.)种质资源开花期关键性状评价

为筛选适宜高寒区种植的草产量高、绿肥特性优的豌豆品种,采用相关性、主成分和聚类分析方法对150份国内外豌豆种质资源花期关键性状进行分析和评价。结果表明:供试豌豆种质资源花期根瘤数量和花干重的变异系数较大,分别为0.65和0.41,其次为叶干重、茎干重和根干重。根瘤数量、茎、叶和花干重间具有显著的相关性。花期主要性状主成分分析发现,前2个主成分累积贡献率为62.76%,分别代表草产量和绿肥特性。系统聚类可将供试豌豆种质资源分为草产量低、草产量高且绿肥特性好、固氮能力最强三大类群。其中,第Ⅱ大类群的保加利亚豌豆、173、秋复5号豌豆、311、A-65豌豆和A-73豌豆等可作为高寒区适宜种植的豌豆种质资源。

Photosynthesis and growth responses of pea Pisum sativum L. under heavy metals stress

The present work aimed to study the physiological effects of cadmium （Cd） and copper （Cu） in pea （Pisum sativum）. Pea plants were exposed to increasing doses of cadmium chloride （CdCl2） and copper chloride （CuCl2） for 20 d. The examined parameters, namely root and shoot lengths, the concentration of photosynthetic pigments and the rate of photosynthesis were affected by the treatments especially with high metals concentrations. The analysis of heavy metals accumulation shows that leaves significantly accumulate cadmium for all the tested concentrations. However, copper was significantly accumulated only with the highest tested dose. This may explain the higher inhibitory effects of cadmium on photosynthesis and growth in pea plants. These results are valuable for understanding the biological consequences of heavy metals contamination particularly in soils devoted to organic agriculture.

Cloning and characterization of 66 kDa streptavidin-binding peptides(SBP)of Pisum sativum L.embryo specific to var.Alaska

The aim of the current research was to clone and to characterize the partial 66 kDa streptavidin-binding peptide(SBP)found in the germinated embryos of Pisum sativum L.var.Alaska.The pea(P.sativum var.Alaska)embryos possess prominent 66 kDa SBPs that gradually disappeared after few hours of germination in germinated embryos,but not in the cotyledons.The total RNA was isolated from embryos of P.sativum but could not be isolated from the cotyledons.The partial nucleotides sequences of 66 kDa SBPs of embryonic stalk(P.sativum var.Alaska)were cloned and identified using pMOSBlue vector.66 kDa(SBP)gene from the embryos of P.sativum var.Alaska possesses 327 bp having an open reading frame(ORF)region in a part of the gene that encoded for 108 amino acids.Alignment showed similarity among 66 kDa SBPs P.sativum var.Alaska,with P.sativum seed biotinylated protein(SBP65)and P.sativum sbp65a mRNA with DNA distance matrix between 0.0094 to 1.2676.MALDI-TOF mass spectrometry analysis of 66 kDa(SBP)proteins showed it had similar short peptides to 19 proteins found in different organisms,especially Convicilin precursor,and the seed biotinylated protein in P.sativum.The alignment results of both nucleotide sequences and amino acid residues either from cloning or MALDI-TOF-MS showed differences with related species,especially P.sativum.No mRNA was found in the cotyledons during seeds germination,which means no metabolic activities and this part may act only as food reservoirs for growing newly embryos.

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.

Subnucleolar Distribution and Transportation of U3 snoRNA in the Nucleolus of Pisum sativum

Processing of pre-rRNA is one of the major events taking place In the nucleolus. U3 snoRNA, an rRNA spliceosomal factor, is suggested to be essential in the first cleavage step of the 5' ETS sequence in the processing of pre-rRNA. Identification of U3 in the nucleolus provides a piece of indirect evidence for pre-rRNA processing site and transportation of processing products. In the present study, subnucleolar distribution of U3 snoRNA in the nucleolus of Pisum sativum L. was studied by in situ hybridization with a U3 snoRNA probe. The results showed that the U3 labeling signals were distributed throughout dense fibrillar components (DFCs) and granular components (GCs), while no signal was found in fibrillar centers (FCs). When treated with actinomycine D (AMD), the labeling signals were decreased. Along with the increase of the AMD treatment time, the labeling signals became fewer and they were found in the distal regions of DFC and GC. Our results indicated that pre-rRNA splicing took place in the regions of DFC and GC, and the transportation of pre-rRNA processing products was from the regions around FCs towards the distal regions.

Gene expression of 49 kDa apyrase,cytoskeletal proteins,ATPase,ADPase and amino acid contents of Pisum sativum(L.)cells germinated in Euryops arabicus(Steud.ex Jaub.&Spach)water extract

The present research reports of quick and marked changes induced by plant extract of Euryops arabicus in the gene expression of 49-kDa apyrases,cytoskeletal proteins,ATPases,ADPase and amount of amino acid of pea(Pisum sativum L.var.Alaska).Pellets of cytoskeletals proteins(27000 xg)were probed with anti-apyrase antibody,biotinylated anti-rat,actin and alpha and beta-tubulin for Western blotting.ATPase and ADPase activities were determined based on the hydrolytic efficacy of adenine triphosphate and adenine diphosphate.By 72 hours,the abundance of apyrases,cytoskeletal proteins and amount of amino acid in pellets of 27000 xg of germinated pea seeds in E.arabicus extracts were sharply increased than those sown in distilled water.All the samples exhibited that the stems had more amount from apyrases,cytoskeletal proteins,amino acids and ATPase and ADPase activities than primary leaves and primary roots that were germinated either on E.arabicus water extract or in distilled water.Based on the enzyme’s capability to hydrolyse nucleotide triphosphate and nucleotide diphosphate as well as the direct association between expression of 49-kDa apyrase and cytoskeletal proteins,E.arabicus water extract had an important effect on plant germinations.

Radiation dose effects on the morphological development of M_(1) generation pea(Pisum sativum)

We irradiated pea seeds with neutrons from a ^(252)Cf source and studied the radiation dose effects on various morphological development parameters during the growth of M_(1) generation peas.We found that in the dose range of 0.51-9.27 Gy,with the increase in neutron-absorbed dose,the morphological development parameters of M_(1) generation peas at the initial seedling stage showed an obvious trend with three fluctuations.With the development of pea,this trend gradually weakened.Further analysis and verification showed that the main trend in the M_(1) generation of pea seeds was an inhibitory effect induced by neutron irradiation and there was a good linear correlation between the inhibitory effect and neutron absorption dose We successfully demonstrated the background removal of mutant plants and defined morphological developmen parameters for peas that match the overall development of plants.Our results will positively impact neutron mutation breeding and automatic agriculture.

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.

Evaluation of Expression Stability of Candidate References Genes among Green and Yellow Pea Cultivars (<i>Pisum sativum</i>L.) Subjected to Abiotic and Biotic Stress

Dry pea (Pisum sativum L.) is grown as human and animal feed throughout the world. Large yield losses in pea due to biotic and abiotic stresses compel an improved understanding of mechanisms of stress tolerance and genetic determinants conditioning these tolerances. The availability of stably expressed reference genes is a prerequisite for examining differential gene expression. The objective of this study was to examine the expression profile of several candidate reference genes across a broad range of commercial pea cultivars. Expression profiles of five candidate reference genes;18s rRNA, actin, TIF, β tubulin-2 and β tubulin-3 were examined. Relative quantifications of candidate reference genes were estimated from control plants, plants after 48 h of cold treatment, and plants 24 and 48 h after inoculation with Sclerotinia sclerotiorum, the causal agent of white mold disease of pea. RT-qPCR was performed on cDNA synthesized from three food grade spring peas, Ariel, Aragorn, and Sterling, and two spring yellow peas, Delta and Universal, which are used as animal feed. Analysis of variance (ANOVA) of CT values demonstrated significant variation between varieties and treatments under cold and disease conditions. The most abundant transcripts among tested reference genes were for 18s rRNA. Stability analysis indicated that TIF and β tubulin-3 genes were the most stably expressed candidate genes under both cold and disease stress and could serve as reference genes across a wide range of pea cultivars.

Screening of field pea (Pisum sativum) selections for their reactions to root-knot nematode (Meloidogyne incognita)

Pot studies were carried out to evaluate the reactions of 23 selections of field pea （Pisum sativum） against Meloidogyne incognita （2000 freshly hatched juveniles （J2）/pot）. Experiment was conducted under greenhouse conditions （（24.7±3） ℃ and （62±7）% RH） and terminated 45 d after inoculation. The roots of all the selections were assessed to determine root-knot indices （RKI） on a 0-4 scale. Out of 23 selections HFP-990713, Pant P-25, and HFP-0129 were resistant; Pant P-2005, NDP-2 and Pant P-42 were tolerant; LFP-305, HFP-8909, HFP-4, HUP-31, HFP-0128, Pant P-31, Pant P-40, LFP-363, and HFP-0118 were moderately resistant; HFP-0110, HUDP-28, HUDP-15, HUDP-27, HUP-30, HUP-2 and HUDP-26 were moderately susceptible; and only Ambika was susceptible to M. incognita. It was observed that reproduction of nematode was favored on tolerant and susceptible cultivars but inhibited on resistant ones. Strong negative correlation was observed between the total fresh and dry plant weights and the root-knot index. The selection Pant P-42 showed highest tolerance among all the selections tested and can be recommended for field trials, whereas, selection Ambika showed highest susceptibility and should be avoided.

Effects of Cadmium Pollution in Soil on Physiological and Biochemical Index of Allium sativum L.

[Objective] The experiment aimed to study the effects of cadmium pollution in soil on physiological and biochemical index of Allium sativum L. and provided reference for the recovery of cadmium pollution in soil. [Method]By setting eleven Cd concentrations from 0.21 to 500 mg/kg in soil and the pot test, ecological corresponding mechanism of plant height, chlorophyll (Chl) content, catalase (CAT) activity and malondialdehyde (MDA) of Allium sativum L. was analyzed. [Result] The plant height had a strong tolerance to cadmium pollution in soil, while the total chlorophyll content and chlorophyll a content had no significant difference compared with control treatment, except Cd concentration was 500 mg/kg. The high Cd concentration would increase the damage to membrane of Allium sativum L. however with the regulation of physiological mechanism, the damage was gradually decreased.[Conclusion] Allium sativum L. had strong eco-physiological adaptability to Cd contaminated soil and it had potential for recovering Cd contaminated soil.

Greenhouse Intercropping of Green Asparagus(Asparagus officinalis Linn) and Coriander(Coriandrum sativum L.)

Greenhouse cultivation in northern China has the characteristic of single mode, and the intercropping mode is not common. Greenhouse intercropping of green asparagus （Asparagus officinalis Linn） and coriander （Coriandrum sativurn L.） not only could increase vegetable kinds, but also could improve cropping index, and the asparagus growers＇ income could also be improved. This paper introduced the intercropping technique of green asparagus and coriander, so as to provide technical support for large-area extension of greenhouse green asparagus cultivation and break the situation of single mode for .qreenhouse cultivation.

大蒜(Allium Sativum L.)对钉螺酯酶同工酶作用的初步研究

为研究在浓度0.05 %大蒜叶水浸液作用下,钉螺的酯酶同工酶谱特征及其变化情况,对采自武汉汉阳的钉螺进行去壳、匀浆,应用聚丙烯酰胺凝胶对匀浆物电泳,进行酯酶同工酶(EST)染色.EST共分离出6～9 条酶带,6 d中相似的酶带有6条,处理不同天数的相应酶带,宽度及染色深浅出现变化,酶带数也出现增减.大蒜水浸液对钉螺的酯酶同工酶有明显影响,开始2天,因受刺激,酶活性(比较对照)增强,随着处理时间延长,酶活性变弱.

Dynamic Study of Microbial Population and Enzyme Activity in Rhizosphere Soil of Allium sativum L.

[Objective] The aim was to investigate the changes in microbial population and soil enzyme activities in rhizosphere soil of two cultivars of Allium sativum L.at different growth stages.[Method]By using white garlic and purple garlic as the experimental materials,the microbial population,and the activities of urease,acid phosphatase and catalase in their rhizosphere soil at different growth stages were measured.[Result]The root exudates of the two garlic cultivars could promote the growth of bacteria,fungi and actinomycetes,and indirectly increase the urease,acid phosphatase and catalase activities in the rhizosphere soil,thereby improving the turnover and circulation of the soil nutrition elements such as nitrogen and phosphorus,and providing a better micro-ecological environment for the later crop.[Conclusion]The study had provided theoretical basis for the ecological research on garlic used as a preceding crop.