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.

Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress(Lepidium sativum L.)

Objective: Garden cress(Lepidium sativum L.) is an important herb in traditional medicine used to improve production of breast milk in women and semen in men. In the present research the authors evaluated its ability to destroy leukemic cancer(Jurkat E6-1) cells, using the alkaloid extract of this plant.Methods: Constituents of the alkaloid extract were analyzed by gas chromatography–mass spectrometry(GC–MS) and their cytotoxicity in leukemic cancer cells and healthy peripheral blood mononuclear cells(PBMCs) was assessed. Cell death via apoptosis was confirmed by DNA laddering, caspase-3 activity,annexin V-fluorescein isothiocyanate and mitochondrial toxicity assays. The specific course of gene activation in treated cells was determined through quantitative polymerase chain reaction(qPCR).Results: GC–MS analysis identified six alkaloids and proto-alkaloids, namely, benzyl isothiocyanate(1),2-ethoxy-4 H-3,1-benzoxazin-4-one(2),(4 R)-2-(2-aminophenyl)-4-phenyloxazoline(3), 5-acetyl-1,2-di hydro-6-methyl-2-oxo-4-phenyl-3-pyridinecarbonitrile(4), benzo[b][1,8]-naphthyridin-5(10 H)-on e,2,4,7-trimethyl(5) and 1,4-diaminoanthraquinone(6), in the alkaloid extract of L. sativum. Of these,compound 1 was previously identified in the seeds of L. sativum. Exposure to the alkaloid extract caused death of Jurkat E6-1 cells, with median lethal concentration(LC50) of 75.25 mg/mL. However, the alkaloid extract also showed a nontoxic and proliferative(1.6-fold) effect in healthy PBMCs. Further experiments performed with Jurkat cells at LC50 and sub-LC50 doses demonstrated DNA fragmentation, activation of caspase-3 and time-dependant phosphatidylserine translocation(apoptosis) from inner to outer cell membranes. Cell toxicity and assessment of adenosine triphosphate level, together with using qPCR to evaluate expression profile of major apoptosis genes, revealed that apoptosis may be induced by disruption in the mitochondrial outer membrane potential, through activation of extrinsic and intrinsic apoptosis pathways in Jurkat cells.Conclusion: The ability of the alkaloid extract of L. sativum seeds to induce apoptosis indicates a potential pharmacological use in cancer chemotherapy. The separation of individual active compounds and further in-depth exploration of the molecular mechanism of apoptosis may lead to novel chemotherapeutic compounds in our future antineoplastic research.

Effects of Lepidium sativum supplementation on growth and gonadotropins secretion in ovariectomized, estrogen-implanted rabbits

Objective: To test the effects of dietary supplementation of Lepidium sativum (LS) seed powder on growth performance and gonadotropins secretion in ovariectomized, estradiol-implanted rabbits. Methods: Ovariectomized, estradiol-implanted Chinchilla rabbits were assigned into four experimental groups: LS seed powder was included into normal rabbit chow at 0% (control), 5% (low), 7% (mid) and 10% (high) w/w. Experimental feed and water were given ad-libitum for 3 weeks. Weekly body weights and daily feed intake of rabbits were recorded. Twenty-one days post-feeding, blood samples were collected at 15-minute interval for 3 h (PeriodⅠ) after which 2.5 μg gonadotropin-releasing hormone (GnRH) was injected intravenously and the sampling continued for another hour (PeriodⅡ). Plasma was harvested and analyzed for luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by radioimmunoassay. Results: Feed intake was significantly (P<0.05) increased in LS-supplemented rabbits. However, the increase in feed intake did not result in significant body weight gain. LS seed supplementation significantly (P<0.001) increased mean plasma LH dose-dependently from the low- to the mid-LS level and then decreased LH at the high-LS level. LS supplementation increased (P<0.001) plasma FSH secretion. Injection of GnRH had no effect on plasma LH, however significantly (P<0.05) decreased overall plasma FSH secretion. Conclusions: LS seed supplementation stimulates feed intake and gonadotropins secretion in rabbits. Gonadotropins effect may be mediated through LS seeds phytosterols through the activation of estrogen receptors thereby producing agonistic effects resulting in LH and FSH secretion. The differential responses of gonadotropins to GnRH in LS-supplemented rabbits suggest differential regulation of the synthesis and secretion of these gonadotropins.

Genetic diversity analysis of Lepidium sativum(Chandrasur) using inter simple sequence repeat(ISSR) markers

Lepidium sativum（commonly known as garden cress） belongs to the family Brassicaceae. It is a fastgrowing erect, annual herbaceous plant. Its seeds possess significant fracture healing, anti-asthmatic, anti-diabetic,hypoglycemic, nephrocurative and nephroprotective activities. In the present study, we assessed the genetic diversity of various genotypes of L. sativum using inter-simple sequence repeat（ISSR） markers. Out of 41 ISSR primers screened, 32 primers showed significant, clear and reproducible bands. A total of 510 amplified bands were obtained using 32 ISSR primers, out of which 422 bands were polymorphic and 88 bands were monomorphic. The percentage of polymorphism was found to be 82. A total of 35 unique alleles ranging insize from 200 to 2,900 bp were observed.Cluster analysis based on unweighted pair-group method,arithmetic mean divided the 18 genotypes into two main clusters, with the first having only HCS-08 genotype of L.sativum and other having all of the other 17 genotypes. The Jaccard similarity coefficient revealed a broad range32–72 % genetic relatedness among the 18 genotypes.

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。

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.

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。

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.

山楂(Crataegus L.)饮料及大蒜(Allium sativum L.)防腐效果研究

以山楂叶、山楂果、蜂蜜、白糖等为原料,用大蒜作为天然防腐剂,研制含天然植物黄酮的健康饮料。通过3×3正交试验确定产品的最佳配方.结果表明,以5％山楂叶汁、50％山楂果汁、8％蜂蜜配制的饮料口感和理化性质较好,并含有保健黄酮;大蒜能有效地防止饮料腐败,有很好的防腐效果。

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.

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.

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.

大蒜MtN3/saliva/SWEET基因家族的全基因组鉴定及表达分析

探究大蒜AsSWEETs基因家族特征和生物学功能,可为大蒜鳞茎发育调控的分子机制提供参考。本研究基于大蒜“二水早”全基因组数据,通过HMM搜索对AsSWEETs成员进行筛选和鉴定,并利用实时荧光定量PCR分析其在不同激素处理下的表达模式。结果表明:在大蒜基因组中共鉴定到35个AsSWEETs基因,均为疏水蛋白,含2~7个不等的跨膜螺旋,除AsSWEET 1、AsSWEET 2位于scaffold外,其余均分布于染色体上,成员间差异显著。综合系统进化和表达模式发现,AsSWEET5、14、21、23、26、28(第III支系,转运蔗糖)可能是参与鳞茎膨大的主要调控基因;AsSWEET 2、30、33(IV分支)的表达量在鳞茎发育不同阶段变化较大,可能参与鳞茎发育过程中的果糖代谢调节。qRT-PCR定量表明,在不同激素处理下,AsSWEET5、11、16呈现出不同的表达模式,预示AsSWEETs家族成员功能存在分化和差异。综上,大蒜AsSWEET s基因的糖转运活性参与了鳞茎的生长发育,有助于解释鳞茎膨大发育过程。

盐胁迫下La^(3+)和Eu^(3+)对小麦质膜能化系统活性的影响

两相法分离纯化小麦(Triticum sativum L)根细胞质膜微囊用于测定La3+和Eu3+对质膜H+-ATPase及氧化还原系统活性的影响。结果显示,200mmol/L NaCl溶液处理显著抑制了小麦幼苗根细胞质膜H+-ATPase活性及NADH氧化速率和Fe(CN)63-还原速率;适宜浓度La3+和Eu3+处理使盐胁迫下小麦活体根细胞质膜H+-ATPase及氧化还原系统活性明显提高;不同浓度稀土离子处理对离体细胞中H+-ATPase活性均表现出了抑制作用。两种稀土离子对活体和离体细胞质膜H+-ATPase具有不同的生理学效应。这些结果将有助于阐明盐胁迫条件下稀土元素提高植物耐盐能力的生理机制。

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.

Plackett-Burman设计和响应面分析法优化香菜中总黄酮的微波提取工艺

为了确定香菜茎叶中总黄酮微波辅助提取最佳工艺条件,首先用Plackett-Burman设计对影响香菜总黄酮得率的因素进行评价,筛选出三个主要影响因素:液料比,微波温度,乙醇体积分数。以香菜总黄酮得率为响应值,采用响应面分析法优化提取条件,确定最佳提取条件为液料比35∶1,微波温度80℃,乙醇体积分数67%。在此条件下黄酮得率的预测值为1.33%,实测值为1.31%,误差为1.30%。表明Plackett-Burman设计结合响应面分析法可以很好的对香菜总黄酮微波提取工艺进行优化。

HPLC内标法测定鲜蒜中蒜氨酸

目的:建立准确、方便的 HPLC 法测定鲜蒜中蒜氨酸。方法:采用微波灭酶,以利巴韦林为内标物,HPLC 紫外检测,测定鲜蒜中蒜氨酸含量。与美国药典蒜氨酸衍生化 HPLC 测定进行比较。结果:同一鲜蒜样品测定蒜氨酸含量,以微波法灭酶大蒜的含量最高,为1.291%(RSD=1.4%),回收率为98.80(±2.0)%;美国药典方法测定结果为1.287%(RSD=3.6%),回收率为96.03(±2.6)%。结论:HPLC 内标法与美国药典方法测定结果一致,操作步骤简化。