Development of flow cytogenetics and physical genome mapping in chickpea (Cicer arietinum L.)

Procedures for flow cytometric analysis and sorting of mitotic chromosomes (flow cytogenetics) have been developed for chickpea (Cicer arietinum). Suspensions of intact chromosomes were prepared from root tips treated to achieve a high degree of metaphase synchrony. The optimal protocol consisted of a treatment of roots with 2mmol/L hydroxyurea for 18h, a 4.5-h recovery in hydroxyurea-free medium, 2h incubation with 10µmol/L oryzalin, and ice-water treatment overnight. This procedure resulted in an average metaphase index of 47%. Synchronized root tips were fixed in 2% formaldehyde for 20min, and chromosome suspensions prepared by mechanical homogenization of fixed root tips. More than 4×10⁵ morphologically intact chromosomes could be isolated from 15 root tips. Flow cytometric analysis of DAPI-stained chromosomes resulted in histograms of relative fluorescence intensity (flow karyotypes) containing eight peaks, representing individual chromosomes and/or groups of chromosomes with a similar relative DNA content. Five peaks could be assigned to individual chromosomes (A, B, C, G, H). The purity of sorted chromosome fractions was high, and chromosomes B and H could be sorted with 100% purity. PCR on flow-sorted chromosome fractions with primers for sequence-tagged microsatellite site (STMS) markers permitted assignment of the genetic linkage group LG8 to the smallest chickpea chromosome H. This study extends the number of legume species for which flow cytogenetics is available, and demonstrates the potential of flow cytogenetics for genome mapping in chickpea.     

Possible antiosteoporotic mechanism of Cicer arietinum extract in ovariectomized rats

Objective: The present study aimed to throw the light on the anti-osteoprotic mechanism of Cicer arietinum extract (CAE) seeds against ovariectomized (OVX) rats. Methods: Seventy female rats were divided into two groups. The first group (14 rats/group) represented normal rats (Sham operated) while the second group (56 rats/group) underwent bilateral ovariectomy (OVX). After one week of recovery from ovariectomy surgery, the second group was randomly subdivided into 4 subgroups (14 rats/ each subgroup). The rats administered orally; distilled water (vehicle) (1^st subgroup), Cicer arietinum extract (CAE) (500 or 1000 mg/kg body weight/day) (2^nd and 3^rd subgroups), alendronate (6.5 mg/kg mg/kg body weight) as a positive control one time/week (4^rh subgroup), daily for 10 weeks. Results: The present study demonstrated that ovariectomy caused significant decrease in bone mineral; density (BMD) and content (BMC), Bone-specific alkaline phosphatase (BALP), calcium (Ca), phosphorus (P), parathyroid hormone (PTH) and calcitonin levels. Furthermore, ovariectomy induced significant elevation of tartrate-resistant acid phosphatase 5b (TRAP 5b) and receptor activator of nuclear factor (NF-kappa β) ligand (RANKL) concentration. Conversely, osteoprotegerin (OPG) and OPG/RANKL ratio were decreased following ovariectomy. The present work suggests that CAE has antiosteoporotic action against ovariectomy effects and its activity may results from its phytochemical and/or phytoestrogen contents. Conclusion: The ongoing study speculates that the CAE exerts its action through regulation of RANK/RANKL/OPG system. As, CAE not only promotes osteoblast differentiation, but also up-regulates OPG and downregulates RANKL secretion in osteoblasts, subsequently prevents bone loss and osteoporosis.     

鹰嘴豆地上部位元素分析及药用价值研究

目的:分析比较鹰嘴豆茎、叶、豆荚中微量元素的含量,为常用维药鹰嘴豆的全面开发、研究提供科学依据.方法:利用电感耦合等离子发射光谱仪分别测定鹰嘴豆茎、叶、豆荚中微量元素的含量,比较各部位微量元素的差异.结果:鹰嘴豆荚、茎、叶中Mg、Ca、Fe、Mn、Zn、Cr等生命必需元素含量较高.结论:从所含蛋白质、维生素、不饱和脂肪酸及微量元素量角度分析,鹰嘴豆有广泛的开发价值.     

鹰嘴豆总皂苷对2型糖尿病大鼠抗氧化作用的影响

目的观察鹰嘴豆总皂苷(TSCA)对2型糖尿病大鼠抗氧化作用的影响。方法以高脂饲料喂养后的雄性Wistar大鼠腹腔注射链脲佐菌素(STZ)诱导2型糖尿病模型,其中两组造模动物分别灌胃给予鹰嘴豆总皂苷100 mg/kg(低剂量组),300 mg/kg(高剂量组)。4周后,测定各组大鼠肝脏、肾脏、肺、骨骼肌等组织中相关抗氧化指标。结果与模型组比较,鹰嘴豆总皂苷低剂量组能够显著升高肾脏中总超氧化物岐化酶(TSOD)水平(P<0.01)、肝脏、肾脏、骨骼肌中总一氧化氮合酶(TNOS)活力(P<0.01)和肺、骨骼肌中谷胱甘肽过氧化物酶(GSH-PX)活性(P<0.01),且能升高骨骼肌中总超氧化物岐化酶(TSOD)水平及过氧化氢酶(CAT)活力(P<0.05),显著降低肝脏、骨骼肌中丙二醛(MDA)的含量(P<0.01);鹰嘴豆总皂苷高剂量组能够显著升高肾脏总超氧化物岐化酶(TSOD)水平(P<0.01)和肝脏、肾脏、骨骼肌中TNOS活力(P<0.01),且能升高骨骼肌总超氧化物岐化酶(TSOD)水平、谷胱甘肽过氧化物酶(GSH-PX)活性、过氧化氢酶(CAT)活力、肾脏过氧化氢酶(CAT)活力(P<0.05),显著降低肝脏MDA的含量(P<0.01)。结论此研究提示鹰嘴豆总皂苷可以改善2型糖尿病大鼠的抗氧化能力。     

鹰嘴豆总皂苷对2型糖尿病大鼠的抗氧化作用

目的:观察鹰嘴豆总皂苷(TSCA)对2型糖尿病大鼠抗氧化作用的影响。方法:以高脂饲料喂养后的雄性Wister大鼠腹腔注射链脲佐菌素(STZ)诱导2型糖尿病模型,造模动物分别灌胃给予鹰嘴豆总皂苷低剂量100 mg/kg、高剂量300 mg/kg,4周后,测定各组大鼠肝脏、肾脏、肺、骨骼肌等组织中相关抗氧化指标。结果:与糖尿病模型组相比,鹰嘴豆总皂苷100 mg/kg、300mg/kg剂量均能升高肝脏、肾脏、骨骼肌、肺组织中的TSOD、GSH-Px活力,升高肝脏、肾脏、骨骼肌中CAT活力,降低MDA含量,降低肺组织中的CAT活力。结论:鹰嘴豆总皂苷能够改善2型糖尿病大鼠的抗氧化能力。     

The Estrogenic Activity of Isoflavones Extracted from Chickpea Cicer arietinum L Sprouts in Vitro

Isoflavones have drawn attention due to their potential therapeutic use. Isoflavones are the important chemical components of the seeds and sprouts of chickpea and higher isoflavones in sprouts than in seeds. However, there have been no previous reports of the estrogenic activity of isoflavones extracted from chickpea Cicer arietinum L sprouts (ICS) in vitro. In this study, which incorporated several in vitro bioassays methods, we systematically evaluated the estrogenic properties of ICS. MTT assay showed that ICS at the low concentration ranges (10^?3–1 mg/L) promoted MCF‐7 cell growth, while at high concentrations, (>1 mg/L) inhibited cell proliferation, indicating ICS worked at a diphasic mechanism. Flow cytometric analysis further calculated the proliferation rate of ICS at low concentration (1 mg/L). ERα/Luc trans‐activation assay and then semi‐quantitative RT‐PCR analysis indicated that ICS at low concentrations induced ERα‐mediated luciferase activity in MCF‐7 cells and promoted the ER downstream target gene pS2 and PR trans‐activation. These effects were inhibited by ICI 182,780, a special antagonist of ER, indicating that an ER‐mediating pathway was involved. Alkaline phosphatase (AP) expression in Ishikawa cells showed that ICS at low concentrations stimulated AP expression. Our current study is the first to demonstrate that ICS has significant estrogenic activity in vitro. ICS may be useful as a supplement to hormone replacement therapy and in dietary supplements. Copyright © 2012 John Wiley & Sons, Ltd.     

响应面法优化鹰嘴豆铁蛋白提取工艺

目的： 优选鹰嘴豆铁蛋白的提取工艺。 方法： 在单因素试验基础上,选择缓冲液pH、料液比、盐析盐摩尔浓度为自变量,干膏量及铁蛋白、总蛋白得率为响应值,根据Box-Behnken原理采用三因素三水平响应面分析法优选鹰嘴豆铁蛋白的提取工艺参数。 结果： 最佳提取工艺条件为缓冲液pH 7.46,料液比1：4,盐析盐摩尔浓度50 mmol·L^-1;铁蛋白得率0.002 556%,与理论预测值0.002 633%偏差较小。 结论： 采用响应面法优选的鹰嘴豆铁蛋白提取工艺稳定合理,为提高铁蛋白得率和植物补铁制剂的开发利用提供参考。     

榆黄蘑提取物对保护肝脏作用的初步探讨

目的探讨榆黄蘑提取物的保肝作用,为开发利用榆黄蘑提供实验基础。方法给动物灌胃给药,腹腔注射环磷酰胺制作小鼠肝损伤模型,测定血清中丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)的活性和丙二醛(MDA)的含量。结果榆黄蘑提取物沉淀部分大剂量(0.8 g/kg/d)和中剂量(0.4 g/kg/d)组对降低肝损伤小鼠血清中ALT和AST的含量有显著效果,上清液小剂量(0.4 g/kg/d)组对降低AST含量也有显著效果;沉淀部分中剂量和上清液小剂量组对MDA含量也有显著降低作用。结论榆黄蘑提取物沉淀部分和上清液均具有明显的保肝作用。     

A PCR-based assay to detect En/Spm-like transposon sequences in plants

Degenerate primers deduced from the TPase region of plant En/Spm-like transposons allowed the amplification of similar sequences from various plant species including sugar beet, wheat and pea. These primers are efficient tools for the detection of this family of transposons in many plant genomes irrespective of sequence knowledge or phenotypic pecularities. An efficient PCR assay was therefore developed for these class II transposons, similar to assays already available for Ty1-copia-, Ty3-gypsy- or LINEs. This approach allowed us not only to show the widespread almost-ubiquitous presence of En/Spm-elements in plant genomes, but also to characterize their genomic organization and chromosomal distribution in the genome of chickpea (Cicer arietinum L.) and its abundance in related Cicer species. This approach can be used for the detection and characterization of endogenous DNA transposable elements in plant species, their complete isolation and evaluation of their use for genome analysis.