香蕉炭疽病拮抗细菌的分离和初步鉴定

对36株从药用植物蛇足石杉中获得内生细菌进行香蕉炭疽病拮抗细菌筛选。采用对峙培养筛选法、碱基序列比对及同源性分析进行研究。结果获得2株对香蕉炭疽病有拮抗作用的菌株,其中JXS1-6和AHL1-1菌株的抑菌带宽达9 mm及4 mm。通过16S rDNA碱基序列比对及同源性分析,JXS1-6和AHL1-1均鉴定为伯克霍尔德属(Burkholderia),属于伯克霍尔德科(Burkholderiaceae),伯克霍尔德目(Burkholderiales),β-变形菌纲(Betaproteobacteria)。     

低温冷害对龙眼成花的影响及抹除荔枝和芒果冷害花穗促进腋芽再生花穗的效果

为探讨早春极端低温对龙眼成花的影响及芒果和荔枝花穗冷害发生时应对措施的效果,在2008年早春低温冷害时,通过抹除芒果和荔枝的冷害顶生花穗,研究该应对措施对促进腋芽再分化花芽并抽生花序的效果,并在低温冷害后,对不同龙眼品种的成花情况进行调查。结果表明:抹除荔枝冷害顶生花穗后能显著促进黑叶、钦州红荔、糯米糍、立夏红腋芽再生花序,平均单株花穗数分别为139、62.5、28和29穗,分别比对照的高119、22.5、25和26穗;而妃子笑、三月红、桂味和禾荔的处理树和对照树之间差异不明显。抹除芒果冷害顶生花穗后,台农1号、贵妃、桂热82号、红象牙和金穗芒的平均单株成花数分别为92、18、131、20.5和18穗,明显高于对照;而凯特芒、桂热120、吉尔、紫花芒和金穗芒处理树和对照树之间差异不显著。龙眼低温冷害后成花较好的有桂明、储良、石硖、小广眼、大乌圆和大广眼,平均单株花穗数分别为88、67、52.7、52、51和50穗;其次是桂香、乌龙岭、东壁、立冬本和早白露,平均单株花穗数分别为39、26、25、23.5和21.5穗。     

Characterization of Colletotrichum lindemuthianum Isolates from the State of Minas Gerais, Brazil

Anthracnose disease of common bean (Phaseolus vulgaris), caused by Colletotrichum lindemuthianum, is responsible for extensive yield losses worldwide. This pathogen is known to vary greatly in its pathogenicity. Control strategies include chemical control and, mainly, the development of resistant cultivars, taking into account the population structure of C. lindemuthianum. The objective of this study was to investigate the pathogenic and genetic diversity and population structure among C. lindemuthianum isolates collected in Minas Gerais state, Brazil. When these isolates were inoculated on 12 differential cultivars, a total of 10 races were identified within a series of 48 isolates collected in Minas Gerais, Brazil. Races 65, 81 and 73 were the most frequent races and occurred in most of the regions. This study also detected race 337, which had not been reported previously in the literature. Random amplified polymorphic DNA (RAPD) analysis performed on the same 48 isolates revealed great genetic diversity, clustering the series into five groups at a maximum similarity value of 89.6%. There was no clear relationship between the loci sampled by RAPD markers and the pathogenic characterization. Analysis of molecular variance showed that 96.06% of the variability was contained within regions and 3.94% among regions, indicating a high exchange of genetic material among the regions of the State. Most of the variability was detected within races (75.24%). The pathogenicity and RAPD assays corroborated the broad genetic diversity of the pathogen and the results have been useful in breeding for resistance to anthracnose.     

Some analytical assays for the determination of bioactivity of exotic fruits

Introduction – The consumption of new exotic fruits, with their high nutritional and sensory value, has significantly increased in the past few years. Among the tropical fruits durian (Durio zibethinus Murr.) is less known than mango (Mangifera indica L.) and avocado (Persea americana). It has been shown that durian, mango and avocado possessed high nutritional and bioactive properties, but these data were determined using different methods. In order to obtain reliable results we investigated samples of durian, mango and avocado of the same stage of ripeness and unified methods were used for determination of the antioxidant potential. As far as we know, no results of such comparative investigation of three tropical fruits (durian, mango and avocado) and the use of such tests for phytochemical control have been published. Objective – Lyophilised durian, mango and avocado samples harvested in 2008 in Thailand and Israel were investigated. Methodology – The contents of crude protein, fat, carbohydrate, dietary fibre, total polyphenols, flavonoids, tannins and flavanols were determined by elemental analysis and UV spectroscopy. The presence of polyphenols (flavonoids and phenolic acids) in the investigated samples was studied by Fourier transform infrared (FT‐IR) spectroscopy and three‐dimensional fluorometry. Four complementary radical scavenging assays were used for antioxidant determination: ferric reducing antioxidant power (FRAP), 2, 2‐azino‐bis (3‐ethyl‐benzothiazoline‐6‐sulfonic acid) diamonium salt (ABTS^?+), 1‐diphenyl‐2‐picrylhydrazyl method (DPPH) and cupric reducing antioxidant capacity (CUPRAC). Chemometrical processing was used for statistical comparison of the fruits. Results – All spectrometric measurements were highly correlated. The contents of total fibre, proteins and fats were significantly higher (p < 0.05) in avocado, and carbohydrates were significantly lower in avocado (p > 0.05) than in the two other fruits. The wavelength numbers of FTIR spectra for three investigated fruits were in the same range (1700–600 cm^?1) as for catechin and gallic acid, used as standards. One main peak could be easily observed at the approximate location of ex/em 275/305 nm and the other one at ex/em 350/430 nm in the methanol polyphenol extracts of investigated fruits in three‐dimensional fluorescence, in contour and cross fluorescence maps. Similarity was found between durian, mango and avocado in polyphenols (9.88 ± 1.0, 12.06 ± 1.3 and 10.69 ± 1.1, mg gallic acid equivalents/g dry weight, d.w.), and in antioxidant assays such as CUPRAC (27.46 ± 2.7, 40.45 ± 4.1 and 36.29 ± 3.7, µM Trolox equivalent (TE)/g d.w.) and FRAP (23.22 ± 2.0, 34.62 ± 3.4 and 18.47 ± 1.9, µM TE/g d.w.), respectively. The multisample median test between all possible pairs of groups is a Tukey–HSD type comparison and denotes the different groups in a case when a pair‐wise test is significant and its q statistical value is greater than the table q parameter. The multisample median test of FRAP values were chosen from the compared fruits triplets as similar or homogenous subsets durian and avocado. Conclusion – Nutritional and bioactive values of durian are comparable with these indices in mango and avocado. These fruits contain high, comparable quantities of basic nutritional and antioxidant compounds, and possess high antioxidant potentials. All fruits show a high level of correlation between the contents of phenolic compounds and the antioxidant potential. The methods used (three‐dimensional fluorescence, FTIR spectroscopy, radical scavenging assays) are suitable for bioactivity determination of these fruits. In order to receive best results, a combination of these fruits has to be included in the diet. The methods used are applicable for bioactivity determination in phytochemical analysis in general. Copyright © 2010 John Wiley & Sons, Ltd.     

Detrital Fruit Processing in a Hawaiian Stream Ecosystem1

In temperate forested streams, fruit from riparian trees is generally a minor and seasonal component of the allo‐chthonous detritus. In contrast, riparian fruit input to tropical streams is often high and continuous. Detrital fruit is abundant in some forested Hawaiian streams compared to other forms of riparian detritus, and rates of leaf litter processing by macroscopic invertebrates are very low. These observations suggested that fruit is an important food resource for detritivores. A microcosm system was used to measure the rates at which two common detrital fruits, guava and mango, were processed by two common detritivores, the prawn Macrobrachium lar and the gastropod Tarebia granifera. Comparisons of fruit weight loss rates normalized by detritivore weight indicated that M, lar processed guavas at significantly higher rates than T. granifera, differences in rates of mango processing by M. lar and T. granifera were not significant. Microcosms containing both M. lar and T. granifera were used to test for interactions between the invertebrates that affected rates of mango processing. No interspecific interactions were detected. A field study was conducted in Kaiwiki Stream, Island of Hawaii, to determine rates of detrital fruit input and export. Detrital fruit was supplied to the study area year‐round, with peaks corresponding to summer and autumn fruiting seasons. Guavas and mangos accounted for 85 percent of the fruit biomass entering the stream and 92 percent of the fruit exported from the stream. Mean daily export rates of guava were 7 percent of input, and export rates of mango were 5 percent of input. These measurements suggested that most of the fruit entering the stream is retained and comprises a substantial food resource for detritivores. Comparisons of the biomass‐specific rates at which M. lar and T. granifera processed mangos and guavas with the rates at which mangos and guavas entered Kaiwiki Stream suggested that these invertebrates can process most of the detrital fruit in the stream.     

Pathogenic and genetic variability among Colletotrichum gloeosporioides isolates from different yam hosts in the agroecological zones in Nigeria

Anthracnose, caused by Colletotrichum gloeosporioides Penz., is the most severe foliar disease of water yam (Dioscorea alata) worldwide. Population genetic analyses can yield useful insights into the evolutionary potential of C. gloeosporioides and thus lead to the development of appropriate disease management strategies. The genetic structure of C. gloeosporioides populations from yam and non‐yam hosts in three agroecological zones of Nigeria was investigated. Microsatellite‐primed polymerase chain reaction (MP‐PCR), virulence phenotyping using five putative D. alata differentials, cross‐inoculation tests, and the presence/absence of a Glomerella teleomorph in yam fields were used to infer the evolutionary potential of C. gloeosporioides on yam. We observed high genotypic diversity (GD = 0.99 to 1.00) for populations from all hosts and agroecological zones, with multiple pathogen genotypes in individual anthracnose lesions. Genetic differentiation was low among pathogen populations from different hosts (G_ST = 0.10, θ = 0.034), and agroecological zones (G_ST = 0.04, θ = 0.018), indicating limited host differentiation and significant gene flow. No evidence was found for the existence of C. gloeosporioides f. sp. alatae reported in previous studies. The fungus was recovered from several non‐yam host species commonly found in yam fields but non‐yam isolates caused only mild to moderate symptoms on yam. Eighteen C. gloeosporioides virulence phenotypes were identified among 217 isolates but there was a weak correlation (r = 0.02, P = 0.40) between virulence phenotype and MP‐PCR haplotype. Consistent with the above findings, we observed for the first time the Glomerella teleomorph on anthracnose‐infected yam plants in Nigeria, indicating that sexual recombination might play an important role in anthracnose epidemics on yam. The implications of these findings for C. gloeosporioides evolutionary potential and anthracnose resistance breeding are discussed.     

Embryogenic mango cultures selected for resistance toColletotrichum gloeosporioides culture filtrate show variation in random amplified polymorphic DNA (RAPD) markers

Summary Genomic DNA isolated from embryogenic cultures of two mango cultivars, ‘Hindi’ and ‘Carabao,’ that had been selected for resistance to the culture filtrate ofColletotrichum gloeosporioides, was analyzed using Randomly Amplified Polymorphic DNA (RAPD).In vitro selection caused changes in RAPD markers in the selected embryogenic cultures with respect to the unchallenged control cultures and the stock plants. The differences involved both the absence and the presence of additional RAPD markers in the resistant lines, although the former was most commonly observed. The absence of differences between the unchallenged control of either cultivar and DNA from the leaves of parent trees confirmed that the changes were not due to prolonged maintenance in liquid cultures.     

Attack of the clones: Population genetics reveals clonality of Colletotrichum lupini,the causal agent of lupin anthracnose

Colletotrichum lupini, the causative agent of lupin anthracnose, affects lupin cultivation worldwide. Understanding its population structure and evolutionary potential is crucial to design successful disease management strategies. The objective of this study was to employ population genetics to investigate the diversity, evolutionary dynamics, and molecular basis of the interaction of this notorious lupin pathogen with its host. A collection of globally representative C. lupini isolates was genotyped through triple digest restriction site-associated DNA sequencing, resulting in a data set of unparalleled resolution. Phylogenetic and structural analysis could distinguish four independent lineages (I–IV). The strong population structure and high overall standardized index of association (r̅_d) indicates that C. lupini reproduces clonally. Different morphologies and virulence patterns on white lupin (Lupinus albus) and Andean lupin (Lupinus mutabilis) were observed between and within clonal lineages. Isolates belonging to lineage II were shown to have a minichromosome that was also partly present in lineage III and IV, but not in lineage I isolates. Variation in the presence of this minichromosome could imply a role in host–pathogen interaction. All four lineages were present in the South American Andes region, which is suggested to be the centre of origin of this species. Only members of lineage II have been found outside South America since the 1990s, indicating it as the current pandemic population. As a seedborne pathogen, C. lupini has mainly spread through infected but symptomless seeds, stressing the importance of phytosanitary measures to prevent future outbreaks of strains that are yet confined to South America.