Physical map of chloroplast DNA of aerial yam,Dioscorea bulbifera L.

Summary A physical map of chloroplast DNA (ctDNA) of aerial yam, Dioscorea bulbifera L. was constructed using three restriction endonucleases, PstI, SalI, and SmaI. In addition, a clone bank of the BamHI-digested fragments were generated, and the locations of most BamHI fragments on the map were also determined. The ctDNA of D. bulbifera was found to be a circular molecule with a total size of ca. 152 kb involving two inverted repeats of ca. 25.5 kb, and small and large single copy regions of ca. 18.5 and 83.4 kb, respectively. The genes for the large subunit of the ribulose 1,5-bisphosphate carboxylase (rbcL) and the ATP-synthase subunits and (atpB/atpE) were mapped.Contribution from the Plant Germ-plasm Institute and the Laboratory of Genetics (No. 504), Faculty of Agriculture, Kyoto University, Japan. The work was supported in part by a Grant-in-Aid (No. 60400005) from the Ministry of Education, Science and Culture, Japan     

In vitro tuberization in white yam (Dioscorea rotundata Poir)

Nodal cuttings of white yam were induced to produce microtubers on a MS-revised medium supplemented with various concentrations of sucrose, 20 mgl^–1 L-cysteine, 0.5 mgl^–1 kinetin and 0.7% agar. The frequency of tuberization was affected by the daylength, which is optimal at 12 and 16 h of light depending on the sucrose concentration. The microtubers were planted in a seed bed and grown to maturity. The importance of in vitro tuberization of yam as a means of international germplasm distribution or exchange as well as for the propagation of planting material is discussed.     

Properties of polyphenol oxidase from tubers of the yam Dioscorea bulbifera

The subunit MW of Dioscorea bulbifera polyphenol oxidase (MW 115 000 ± 2000) determined by SDS-PAGE is ca. 31 000 indicating that the enzyme is an oligomeric protein with four subunits. K_i values of various inhibitors and their modes of inhibition have been determined with catechol and pyrogallol as substrates. p-Nitrophenol, p-cresol, quinoline and resorcinol are competitive inhibitors of catechol binding while only orcinol and p-nitrophenol behave in the same way towards pyrogallol as substrate. From the effect of pH on V_max, groups with pK values ca. 4.7 and 6.8 have been identified to be involved in catalytic activity. The Arrhenius activation energy (E_a) at pH 4.0 is 8.9 kcal/mol between 40–65°. At pH 7.0, the value is 22.1 kcal/mol between 40 and 60°. The enthalpies (ΔH) at pH 4.0 and pH 7.0 are 2.3 kcal/mol and 32.4 kcal/mol respectively. The results are discussed considering the conformational changes of the enzyme during substrate binding.     

Purification and some properties of polyphenol oxidase from the yam tubers, Dioscorea bulbifera

In a comparison of the polyphenol oxidase activity of various species of yam tubers the greatest enzyme activity was found in D. bulbifera. The enzyme was purified from acetone powder extracts of this plant. Ammonium sulphate fractionation, followed by ion exchange chromatography and gel filtration gave 22-fold purification. The final product gave a single band on polyacrylamide disc gel electrophoresis. The purified enzyme showed activity towards catechol, pyrogallol and dl-β-3,4-dihydroxyphenylalanine (dl-DOPA) and had a MW 115000 ± 2000. It was characterized by response to various inhibitors. β-Mercaptoethanol, dithioerythritol, l-cysteine, sodium metabisulphite and KCN inhibited strongly.     

辅色剂对紫淮山花色苷稳定性的影响及其热降解动力学研究

为探究谷胱甘肽和没食子酸对紫淮山花色苷的辅色作用,本文研究了谷胱甘肽和没食子酸对紫淮山花色苷降解率、热稳定性及色差的影响。试验表明,谷胱甘肽和没食子酸能有效抑制花色苷的降解,且最佳添加量分别为0.03%和0.2%。在此添加量条件下,紫淮山花色苷在50、70、90℃水浴中的热降解均符合一级降解反应动力学规律。添加谷胱甘肽和没食子酸的紫淮山花色苷降解速率常数(k)小于对照组,半衰期(t 1/2)和活化能(Ea)高于对照组,说明谷胱甘肽和没食子酸能够增强花色苷的热稳定性。色差测定结果表明,经谷胱甘肽和没食子酸辅色后的紫淮山花色苷,其明度指数(L*)和色品指数(a*、b*)较对照组变化缓慢,颜色的稳定性增强。     

Serological detection of yam viruses in farmers' fields in Ogun State,Nigeria

A field survey was conducted in eight local government areas (LGA) of Ogun state, Nigeria to assess the incidence of viral diseases of yams in the areas. Leaf samples were collected from 90 yam plants which were either symptomatic or asymptomatic. These were bulked into 45 during serological tests and the viruses indexed include yam mosaic virus (YMV); Dioscorea alata bacilliform virus (DaBV) and cucumber mosaic virus (CMV). DaBV was the most prevalent virus on the field with incidence of 48.9% (22/45) followed by YMV which occurred in 42.2% (19/45). CMV had the lowest percentage of incidence; 2.2% (1/45). Of all the LGAs visited, Abeokuta north and Abeokuta south had the highest incidence of YMV and DaBV, respectively. Mixed virus infections were also detected.     

Evaluation of neem (Azadirachta indica) seed and ginger (Zingiber officinale) as potential control agents of yam (Dioscorea rotundata Poir.) tuber rot fungi

Yam is an important crop which serves as a source of income to small-holder farmers as well as a foreign exchange earner. Among the constraints faced by yam farmers are pests and diseases, especially deterioration caused by microbes during storage. Since over dependence on pesticides is being discouraged, neem seed and ginger extracts were evaluated as potential control agents against rot-causing fungi. The study was conducted in the Spanish laboratory, at the Faculty of Agriculture, University for Development Studies, Nyankpala Campus. Isolations were made from rotted yam tubers sampled from the Tamale Central market with potato dextrose agar (PDA) as the growth medium. Growth inhibition of the isolates was determined by growing pure cultures on PDA plates amended with 2?ml each of ethanol and aqueous extracts of neem seed and ginger as well as carbendazim. A pathogenicity test proved that Aspergillus flavus, A. niger, A. ochraceus and Penicillium digitatum isolated from the rotted tubers were responsible for the rot. A. niger had a significantly higher (p?<?0.05) occurrence (40%) than the others. Growth inhibitions by carbendazim and ethanol extracts of neem seed and ginger were comparable but significantly higher (p?<?0.05) than the aqueous extracts of neem seed and ginger as well as the control. However, the aqueous extracts of neem seed and ginger had a significantly higher (p?<?0.05) inhibition than the control. For instance, growth inhibition of A. niger by carbendazim, ethanol neem seed and ginger extracts were 70.5, 69.6 and 65.5%, respectively, while inhibition by aqueous neem seed and ginger extracts were 33.9 and 24.8%, respectively. Since aqueous extracts of neem seed and ginger significantly inhibited (p?<?0.05) growth of the rot-causing fungi, they can be used as surface protectants of stored tubers.     

Mycoflora of tuber surface of white yam (Dioscorea rotundata poir) and postharvest control of pathogens with Bacillus subtilis

Bacillus subtilis (Enrenberg) Cohn was investigated for its antagonistic properties against surface mycoflora of yam (Dioscorea rotundata Poir) tubers in storage. Yam tubers inoculated with a spore suspension of B. subtilis in potato dextrose broth using a knapsack sprayer showed a drastic reduction in the range and number of mycoflora, including pathogens of the tuber surface in contrast to the control tubers, during the five-month storage period in a traditional yam barn. However, B. subtilis maintained a high frequency of occurrence during the same period. Botryodiploidia theobromae Pat, Fusarium moniliforme Wollen and Reink., Penicillium sclerotigenum Yamamoto, and Rhizoctonia sp. were displaced completely on the treated tubers. The antagonism of B. subtilis was so effective that the normal tuber surface mycoflora was greatly reduced throughout the storage period of five months by a simple initial application of the antagonist.This revised version was published online in October 2005 with corrections to the Cover Date.     

Isolation of high quality RNA from seeds and tubers of the Mexican yam bean (Pachyrhizus erosus)

Pachyrhizus erosus is a tuberous legume native to Central America that has great potential for development as a food crop. It produces both protein rich grain and starch filled tubers. There are two major limitations to its dietary use, the high levels of rotenone found in the grains and the low starch content of the tubers, both of which must be addressed, for development of the crop. The low variability of the existing gene pool of the genus limits the use of conventional plant breeding to address these problems. Genetic engineering technology is, therefore, being adopted. For this purpose, an efficient means of RNA isolation from yam bean tissues was developed. The quality of RNA obtained by this method was tested byin vitro translation and was sufficient for use in RT-PCR.