冬瓜和节瓜DNA提取及RAPD反应体系优化

以8份冬瓜和节瓜为材料,采用改良CTAB法提取基因组DNA,采用正交试验设计,对冬瓜和节瓜RAPD条件进行了优化,建立了最佳反应体系:25μL反应体系中含1×buffer,模板DNA、Mg2+、dNTPs、引物和Taq酶的浓度分别为20 ng、2.0mmol/L、0.24 mmol/L、0.3μmol/L和1.0 U。PCR扩增程序为:94℃预变性5 min;94℃变性45 s,36.9℃退火45 s,72℃延伸1.5min,共40个循环;72℃延伸10 min,12℃保存。     

Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Reconstructing the origins and dispersal of the Polynesian bottle gourd (Lagenaria siceraria)

The origin of the Polynesian bottle gourd (Lagenaria siceraria), an important crop species in prehistoric Polynesia, has remained elusive. Most recently, a South American origin has been favored as the bottle gourd could have been introduced from this continent with the sweet potato by Polynesian voyagers around A.D. 1,000. To test the hypothesis of an American origin for the Polynesian bottle gourd, we developed seven markers specific to bottle gourd (two chloroplast and five nuclear). The nuclear markers were developed using a new technique where polymorphic inter simple sequence repeat (ISSR) markers are converted into single-locus polymerase chain reaction and sequencing markers--an approach that will be useful for developing markers in other taxa. All seven markers were sequenced in 36 cultivars of bottle gourd from Asia, the Americas, and Polynesia. The results support a dual origin for the Polynesian bottle gourd: the chloroplast markers are exclusively of Asian origin, but the nuclear markers show alleles originating in both the Americas and Asia. Because hybridization of Polynesian bottle gourds with post-European introductions cannot be excluded, ancient DNA from archaeological material will be useful for further elucidating the prehistoric movements of this species in Polynesia. This work has implications not only for the dispersal of the Polynesian bottle gourd but also for the domestication and dispersal of the species as a whole.     

节瓜、冬瓜重要性状资源表型多样性及亲缘关系的SRAP分析

以冬瓜、节瓜38份种质资源为材料,主要对萌芽率、单果重、瓜长、瓜宽、瓜皮色、瓜型、星点多少、果肉厚和果肉色共计9个性状的表型性状进行了研究,供试材料9个性状的表型变异系数在9.12%~123.13%之间,尤以单果重的变异系数最大;遗传多样性指数(H’)在0.122~1.859之间。SRAP结果显示:利用筛选出的20对引物对38份节瓜、冬瓜材料进行扩增,共扩增出多态性条带323条,遗传相似系数在0.95~0.99之间,节瓜和冬瓜的遗传背景非常窄,冬瓜和黄毛型节瓜亲缘关系近于普通类型节瓜。本研究还解决了部分材料同名异物的问题,可为冬瓜和节瓜育种提供有价值的参考。     

华南瓜类疫霉的RAPD遗传多样性分析

为了解来自广东和广西的瓜类疫霉的遗传多样性,利用从180条RAPD引物中所筛选出的多态扩增性强、重复性好的12条引物,对分离自两省区的96株瓜类疫霉进行了全基因组DNA遗传多样性分析和指纹图谱构建。通过对供试菌株的RAPD-PCR扩增,共获得135条DNA标记谱带,其中124条为多态性谱带,多态检测率为91.9%。利用NTSYSpc Version2.1软件对供试菌株间的遗传距离进行聚类分析并构建系统树,以遗传相似系数0.81为阈值,将96个供试菌株划分为12个RAPD群,多数分离物之间遗传相似性较低,在DNA水平上存在显著的遗传变异,具有较丰富的遗传多样性。不同地区间菌株的遗传分化程度不同,分离自黄瓜的菌株遗传分化明显高于分离自冬瓜的菌株。RAPD群与菌株地理来源、分离寄主、致病力、交配型及甲霜灵抗性均无明显的相关性。     

Heavy metals uptake by vegetables growing in sewage irrigated soil: relationship with heavy metal fractionation in soil

Abstract

Concentrations of the heavy metals Cd, Ni, Cr, Zn, Mn, Co and Cu in soils and vegetable samples (i.e. green chili and gourd) taken from six vegetable fields in the vicinity of Lahore, Pakistan were measured. These soils have been irrigated for a long time with untreated sewage effluents. A control site was selected that has history of fresh canal water irrigation. The sequential extraction procedure developed by Tessier was adopted to demarcate five metal fractions: exchangeable, acid soluble, reducible, oxidizable and residual. The extractants and digests were analysed by atomic absorption spectrometry. The fractionation procedure showed that all the metals were dominant (>50%) in the residual phase in control as well as in waste waster irrigated soils. The concentrations of all the metals in edible parts of the vegetables collected from wastewater irrigated soils, were above critical levels. The total metal concentrations in wastewater irrigated soils followed the order Mn> Co> Zn> Cr> Ni> Cu> Cd, while in control soils the ord er was Mn> Zn> Ni> Co> Cr> Cu> Cd. In order to understand the uptake of metals from soil to vegetables, correlation analyses were performed between metal concentrations in different fractions of soil and their concentrations in vegetables. Correlation analysis was performed at 95% and 99% confidence level. The meaningful significant negative correlation was observed between Cd_F5–Cd_chili,Cr_Total–Cr_gourd,Mn_F5–Mn_Chili,Gourd, Co_F5, _Total–Co_chili, Cu_Total–Cu_Chili which indicate the non availability of total content and residual fraction to studied vegetables. The positive correlation was observed between Cd_Fl-Cd_ChiU, Cd_Fl–Cd_gourd, Ni_{Fi, F3}–Ni_Gourd, Cr_{Fi, F3}, _F4–Cr_ChiU, Cr_F2–Cr_Gourd, Zn_{Fi, F2}–Zn_Gourd, Mn_F3–Mn_Chili, Mn_Fi–Mn_Gourd, Co_F2–Co_Chili, Cu_Fi–Cu_Chili. The positive correlation, especially for the first (exchangeable) second (acid soluble) and for the third (reducible) extraction steps, was obtained which indicate the bioavailability of these metal fractions to plant.