Inheritance of resistance in water yam (Dioscorea alata) to anthracnose (Colletotrichum gloeosporioides)

Colletotrichum gloeosporioides causes anthracnose, the most severe foliar disease of field-grown water yam (Dioscorea alata). The inheritance of resistance to a moderately virulent (FGS) strain of the pathogen was investigated in crosses between tetraploid D. alata genotypes: TDa 95/00328 (resistant)×TDa 95–310 (susceptible) (cross A), and TDa 85/00257 (resistant)×TDa 92–2 (susceptible) (cross B). Segregation of F₁ progeny fitted genetic ratios of 3:1, 5:1 (crosses A and B) and 7:1 (cross A) resistant:susceptible when inoculated with the FGS strain, indicating that resistance is dominantly inherited and suggesting that more than one gene controls the inheritance of resistance to this strain in the accessions studied. When parental and progeny lines of cross A were inoculated with an aggressive (SGG) strain of the pathogen, all plants expressed a susceptible phenotype, indicating strain-specific resistance in TDa 95/00328. Screening of 20 cultivars/landraces confirmed the high susceptibility of D. alata accessions to the SGG strain and revealed the presence of apparent strain non-specific resistance in TDa 85/00257. TDa 85/00257 and TDa 87/01091 which were resistant to the SGG strain, will be useful both as sources of resistance and in the development of a host differential series for D. alata. Received: 15 May 2000 / Accepted: 18 October 2000     

Biological control of anthracnose (Colletotrichum gloeosporioides) in yam by Streptomyces sp.MJM5763

Aim: To find a suitable biocontrol agent for yam anthracnose caused by Colletotrichum gloeosporioides. Methods and Results: An actinobacterial strain, MJM5763, showing strong antifungal activity, multiple biocontrol and plant growth‐promoting traits was isolated from a yam cultivation field in Yeoju, South Korea. Based on morphological and physiological characteristics and analysis of the 16S rDNA sequence, strain MJM5763 was identified as a novel strain of Streptomyces and was designated as Streptomyces sp. MJM5763. Treatment with MJM5763 and the crude culture filtrate extract (CCFE) was effective in suppressing anthracnose in detached yam leaves in vitro and reduced incidence and severity of anthracnose in yam plants under greenhouse conditions. The CCFE treatment was the most effective of all the treatments and reduced the anthracnose severity by 85–88% and the incidence by 79–81%, 90 days after inoculation with the pathogen. CCFE treatment was also effective under field conditions and showed a reduction of 86 and 75% of anthracnose severity and incidence, respectively. Conclusion: Streptomyces sp. strain MJM5763 was effective in biocontrolling anthracnose in yam caused by C. gloeosporioides. Significance and Impact of the Study: Streptomyces sp. MJM5763 is a potential alternative to chemical fungicides for reducing yield losses to anthracnose in yam.     

Assessment of potential antagonists for anthracnose (Colletotrichum gloeosporioides) disease of mango (Mangifera indica L.) in North Western Ethiopia (Pawe)

Mango (Mangifera indica L.) is considered as one of the most popular fruits among millions of people in the tropical area and increasingly in the developed countries. Anthracnose, caused by the fungus Colletotrichum gloeosporioides, is the most important pre- and post-harvest disease of mango. The objective of this research was to evaluate the prevalence of different promising antagonistic Trichoderma and Bacillus spp. on phyloplane of mango in Ethiopia and to evaluate their antagonistic potential against the pathogen. A total of 19 mango fields were surveyed and anthracnose affected all fields. Culture studies on potato dextrose agar for evaluation of antibiosis activity of Trichoderma spp. and Bacillus spp. revealed that they have inhibitory and lytic effect on C. gloeosporioides, which is an indication of their potential biocontrol agent for management of mango anthracnose as an alternative to chemical control. Significant differences (p?<?0.05) were observed among Bacillus isolates in causing lysis of pathogen mycelium, when inoculated on actively growing colony of C. gloeosporioides. Maximum reduction in growth rate of pathogen was observed with Bacillus spp. (B50), which restricted the growth to 2.7?mm compared to 8.3?mm in the control with 67.5% efficacies. There were similar effects (p?<?0.05) among Trichoderma spp. in formation of inhibition zones and lysis by varying degrees up to 59.7% efficacies in reducing linear growth of the pathogen in dual culture.     

PCR-based detection and characterization of the fungal pathogens Colletotrichum gloeosporioides and Colletotrichum capsici causing anthracnose in papaya (Carica papaya l.) in the Yucatan peninsula

Colletotrichum gloeosporioides is the common causal agent of anthracnose in papaya (Carica papaya L.) fruits, and infection by this fungal pathogen results in severe post-harvest losses. In the Yucatán peninsula (Mexico) a different Colletotrichum species was isolated from papaya fruits with atypical anthracnose lesions. The DNAs from a variety of Colletotrichum isolates producing typical and atypical lesions, respectively, were amplified by PCR with C.gloeosporioides-specific primers. All isolates from typical anthracnose lesions yielded a 450 bp PCR product, but DNAs from isolates with atypical lesions failed to produce an amplification product. For further characterization, the rDNA 5.8S-ITS region was amplified by PCR and processed for sequencing and RFLP analysis, respectively, to verify the identity of the papaya anthracnose pathogens. The results revealed unequivocally the existence of two Colletotrichum species causing anthracnose lesions on papaya fruits: C. gloeosporioides and C. capsici. PCR-RFLP using the restriction endonuclease MspI reliably reproduced restriction patterns specific for C. capsici or C. gloeosporioides. The generation of RFLP patterns by MspI (or AluI or RsaI) is a rapid, accurate, and unequivocal method for the detection and differentiation of these two Colletotrichum species.     

Molecular diagnosis of Colletotrichum gloeosporioides causing Anthracnose/Dieback disease in Greater Yam (Dioscorea alata L.)

The Greater Yam (Dioscorea alata L.), a significant tropical tuber crop is highly affected by the anthracnose/dieback disease caused by Colletotrichum gloeosporioides, which greatly reduces the yield as well as market acceptability of the tubers. The different methods that could be used for proper identification of the pathogen by PCR were investigated. The studies indicate that species specific polymerase chain reaction assay based on highly conserved regions in ITS in the genome of the pathogen can be the best strategy for detection of this pathogen at species level. The use of genus specific primers was also successful in detection of Colletotrichum spp. The cloning and sequencing of evolutionarily conserved regions such as ITS, PelB and paralleling them with the available sequences in NCBI database is also costly but reliable approach. The various methods are elaborately tested and their use in diagnosis is discussed in detail.     

Physiological characterisation of Colletotrichum gloeosporioides,the incitant of anthracnose disease of noni in India

Ten isolates of Colletotrichum gloeosporioides were collected from different noni growing areas of Tamil Nadu, Karnataka and Kerala in India and their pathogenicity was proved under glass house conditions. Effect of different pH levels, temperature, light intensity and media were tested against the growth of C. gloeosporioides under in vitro. The results indicated that the growth of C. gloeosporioides was maximum in pH range of 6.50–7.00 and temperature range of 25–30°C. Exposure of the fungus to alternate cycles of 12 h light and 12 h darkness resulted in the maximum mycelial growth of C. gloeosporioides compared to the 24 h exposure to continuous light and 24 h exposure to continuous dark. Among the different media tested, host leaf extract medium supported significantly the maximum growth of all the 10 isolates of C. gloeosporioides followed by potato dextrose agar. Further, the strains were found to vary morphologically between the isolates under the study.     

Sensitivity of mango anthracnose pathogen, Colletotrichum gloeosporioides, to the fungicide prochloraz in Taiwan

In order to monitor the sensitivity of the mango anthracnose fungus, Colletotricthum gloeosporioides, to the eradicative imidazole fungicide prochloraz, a total of 43 mango orchards were surveyed throughout the Tainan area, covering a 4000 ha region of mango plantations. These orchards were recognized as having undergone higher prochloraz application. A subpopulation, 55 isolates in total, collected from Wufeng, Taichung, served as a baseline population since no fungicide was ever used in this mango plantation. A total of 545 isolates were surveyed, and it was found that the IC50s (50% inhibitory concentration) fell within a range of 0.009-0.1554 microg/ml. No significant resistance was found in the field even with higher frequency of prochloraz application. One orchard (Yujing (Wu)) located in the Yujing area known had a higher frequency of prochloraz application, the IC50s were between 0.0204 - 0.1554 microg/ml. The average IC50 was 0. 0766 microg/ml, which was about five times higher than the baseline population (0.015 microg/ml). A significant t test indicated that these two sub-populations were different at p = 0.01. The results indicated that the dose-response of C. gloeosporioides to prochloraz shifted slightly toward higher IC50 over time. A further survey using 10 microg/ml as the threshold dosage was conducted, and the results of 1375 isolates collected throughout this region showed that no isolate could survive at this dosage. Knowing that the registered dosage for field use is 83.3 microg/ml, based on above results, the author concluded that there was no sign of prochloraz resistance in mango plantations 13 years after prochloraz registration in Taiwan.     

Compositional and enzymatic changes in guava (Psidium guajava L.) fruits during ripening

Changes in chemical composition and hydrolytic enzyme activities in guava fruits cv. Lucknow-49 have been reported at four different stages of maturity, viz., mature green (MG), color turning (CT), ripe (R) and over ripe (OR). Chlorophyll content decreased, while carotenoid content increased with advancement of ripening. Starch content decreased with concomitant increase in alcohol soluble sugars. The cell wall constituents viz., cellulose, hemicellulose, and lignin decreased up to R stage, while the pectin content decreased throughout up to OR stage. Among the cell wall hydrolyzing enzymes, polygalacturonase (PG) and cellulase exhibited progressive increase in activity throughout ripening, while pectin methyl esterase (PME) activity increased up to CT stage and then decreased up to OR stage. The maximum increase in the activities of cell wall hydrolysing enzymes was observed between MG and CT stages. The activities of starch hydrolyzing enzymes, α-amylase and β-amylase decreased significantly with advancement of ripening. These changes in the activities of hydrolyzing enzymes could be considered good indicators of ripening in guava.     

蓝光诱导的胶孢炭疽菌(Colletotrichum gloeosporioides)类胡萝卜素积累

胶孢炭疽菌(Colletotrichumgloeosporioides)为一种丝状真菌,蓝光照射可诱导类胡萝卜素的积累。光镜下观察表明,蓝光可诱导胶孢炭疽菌菌丝积累色素颗粒,而黑暗和红光处理却无此现象。类胡萝卜素的积累受蓝光光照强度的影响。28℃且蓝光为6.5μmol.m-2.s-1时,类胡萝卜素积累量可随光照时间延长呈增长趋势,在第5天达到最高峰为71.8μg/g FW,随后含量下降。此外,胶孢炭疽菌在黑暗中预培养的时间也影响蓝光的诱导反应。     

Isolation of antimicrobial compounds from guava (Psidium guajava L.) and their structural elucidation

Four antibacterial compounds were isolated from leaves of guava (Psidium guajava L.), and the structures of these compounds were established on the basis of chemical and spectroscopic evidence. Two new flavonoid glycosides, morin-3-O-alpha-L-lyxopyranoside and morin-3-O-alpha-L-arabopyranoside, and two known flavonoids, guaijavarin and quercetin, were identified. The minimum inhibition concentration of morin-3-O-alpha-L-lyxopyranoside and morin-3-O-alpha-L-arabopyranoside was 200 microg/ml for each against Salmonella enteritidis, and 250 microg/ml and 300 microg/ml against Bacillus cereus, respectively.     

Transferability of simple sequence repeat (SSR) markers developed in guava (Psidium guajava L.) to four Myrtaceae species

Present study demonstrated the cross-genera transferability of 23 simple sequence repeat (SSR) primer pairs developed for guava (Psidium guajava L.) to four new targets, two species of eucalypts (Eucalyptus citriodora, Eucalyptus camaldulensis), bottlebrush (Callistemon lanceolatus) and clove (Syzygium aromaticum), belonging to the family Myrtaceae and subfamily Myrtoideae. Off the 23 SSR loci assayed, 18 (78.2 %) gave cross-amplification in E. citriodora, 14 (60.8 %) in E. camaldulensis and 17-17 (73.9 %) in C. lanceolatus and S. aromaticum. Eight primer pairs were found to be transferable to all four species. The number of alleles detected at each locus ranged from one to nine, with an average of 4.8, 2.6, 4.5 and 4.6 alleles in E. citriodora, E. camaldulensis, C. lanceolatus and S. aromaticum, respectively. The high levels of cross-genera transferability of guava SSRs may be applicable for the analysis of intra- and inter specific genetic diversity of target species, especially in E. citriodora, C. lanceolatus and S. aromaticum, for which till date no information about EST-derived as well as genomic SSR is available.     

Biotechnological advances in guava (Psidium guajava L.): recent developments and prospects for further research

Guava (Psidium guajava L.), an important fruit crop of several tropical and sub-tropical countries, is facing several agronomic and horticultural problems such as susceptibility to many pathogens, particularly guava wilting caused by Fusarium oxysporium psidii, low fruit growth, short shelf life of fruits, high seed content, and stress sensitivity. Conventional breeding techniques have limited scope in improvement of guava owing to long juvenile period, self incompatibility, and heterozygous nature. Conventional propagation methods, i.e., cutting, grafting or stool layering, for improvement of guava already exist, but the long juvenile period has made them time consuming and cumbersome. Several biotechnological approaches such as genetic transformation may be effective practical solutions for such problems and improvement of guava. The improvement of fruit trees through genetic transformation requires an efficient regeneration system. During the past 2–3 decades, different approaches have been made for in vitro propagation of guava. An overview on the in vitro regeneration of guava via organogenesis, somatic embryogenesis, and synthetic seeds is presented. Organogenesis in several different genotypes through various explant selection from mature tree and seedling plants has been achieved. Factors affecting somatic embryogenesis in guava have been reviewed. Production of synthetic seeds using embryogenic propagules, i.e., somatic embryos and non-embryogenic vegetative propagules, i.e., shoot tips and nodal segments have also been achieved. Development of synthetic seed in guava may be applicable for propagation, short-term storage, and germplasm exchange, and distribution. An initial attempt for genetic transformation has also been reported. The purpose of this review is to focus upon the current information on in vitro propagation and biotechnological advances made in guava.     

Induction of phenolic compounds in Hypericum perforatum L. cells by Colletotrichum gloeosporioides elicitation

Changes in phenolic metabolism after elicitation with Colletotrichum gloeosporioides (CG) has been studied in Hypericum perforatum L. (HP) cell suspension cultures. Soluble phenolics were analysed by HPLC-DAD and HPLC-DAD-MS/MS. HP cultures elicited with the CG elicitor showed a significant increase in xanthone accumulation. Xanthone accumulation increased twelve fold when the cells were primed with methyl-jasmonate (MeJ) or salicylic acid (SA), before elicitation. HP cultures exposed only to MeJ produced a set of flavonoids, the flavones which represent a substantial part (approx. 40%) of the total flavonoids accumulated in these cells. The possible importance of xanthones as a component of defence mechanism of HP against biotic stress is discussed.     

In vitro shoot proliferation and propagation of guava (Psidium guajava L.) from germinated seedlings

Guava seeds were germinated on Murashige and Skoog (MS) medium with or without 8.8 μM benzyladenine (BA). BA increased the rate of germination and the number of lateral shoots (3.4 vs 1.2 per seedling). Stem nodes from these lateral shoots were cultured on proliferation media with 4.4 μM BA, and multiple shoots (3.5) were formed within 4 weeks of culture. Increasing the concentration of BA or the addition of naphthaleneacetic acid (NAA) did not affect shoot formation. Shoots produced from explants and lateral shoots from germinated seedlings were rooted in media containing activated charcoal (AC) or 9.8 μM indolebutyric acid (IBA). Shoots rooted with IBA had a higher rooting percentage (100% vs 75%) and a greater number of roots (5.5 vs 3.2) but the shoots were shorter (2.6 vs 3.4 cm) than when rooted in AC, and they required an additional 4 weeks of culture in media with AC to achieve shoot elongation. About 80% of the shoots with roots survived in the glasshouse and produced normal phenotypic plants.     

Isolation of defense-related genes differentially expressed in the resistance interaction between pepper fruits and the anthracnose fungus Colletotrichum gloeosporioides

Unripe mature green fruits of pepper (Capsicum annuum) are susceptible to Colletotrichum gloeosporioides, whereas ripe red fruits are not. We established this pepper-C. gloeosporioides interaction as a model system to study the fungal resistance that develops during ripening of nonclimacteric fruit. Histochemical examination of transverse sections suggested that fungal invasion 24 h after inoculation (HAI) and colonization 48 HAI are critical events that differentiate between resistant and susceptible interactions. Based on this observation, we used messenger RNA differential display to isolate defense-related genes differentially expressed at 24 and 48 HAI. RNA gel blot analysis showed that six out of eighty cloned cDNAs were differentially expressed after infection of ripe fruit. The proteins encoded by these six clones, ddP1, ddP3, ddP4, ddP6, ddP13, and ddP47, showed significant homology to aldehyde dehydrogenase, P23 protein, NP24 protein, cytochrome P450 protein, esterase, and MADS-box protein, respectively, and may be involved in the resistance of ripe fruit to C. gloeosporioides infection.     

Some epidemiological aspects of post-bloom fruit drop disease (Colletotrichum gloeosporioides) in citrus

Fluctuations in the incidence and amount of post-bloom fruit drop disease of citrus caused by Colletotrichum gloeosporioides in Belize prevent economic disease control. During the cooler drier months of the year when blossom infection is common there are variations in the incidence of rainfall and associated climatic parameters, and in the pattern of flowering. Large amounts of disease develop when periods of rain followed by prolonged wetness occur during peak blossoming periods. Blossoms are most susceptible during the open flower stage and infection of terminal flowers invariably results in infection of all other flowers on the spike. Disease incidence is greater in the lower parts of the trees, but flowering is greater in the upper regions. Large numbers of Colletotrichum spores are produced during wet conditions from apparently healthy leaves and from diseased flowers, but these rapidly lose viability when dried. Few spores are produced from old persistent calices (buttons). Although spores from leaves were a less potent inoculum source than those from flowers, they could provide the initial inoculum to commence flower infection when blossoming starts.     

Metabolic analysis of guava (Psidium guajava L.) fruits at different ripening stages using different data-processing approaches

Gas chromatography coupled with time-of-flight mass spectrometry and principal component analysis were used to obtain the metabolite profiles of guava (Psidium guajava) fruits. Results with two types of data-processing software, ChromaTOF and AMDIS, were compared to explain the differences between the samples. There were some differences in score and loading plot patterns of PCA as well as in the composition of the metabolites. However, little difference was observed in the type of metabolites detected and identified using either type of software. Both the flesh and peel of premature and mature white guava fruits were compared for the analysis of the metabolite profiles. Malic acid, aspartic acid, and glucose were the major metabolites distinguishing the different parts of guava fruits in the PCA loading plot. In addition, the metabolic profiles of the fruits revealed significant changes in some metabolites during ripening. The major components contributing to the separation were serine, citric acid, fructose, sucrose, and some unknowns. In particular, sucrose, fructose, serine and citric acid were related to the ripening of guava fruits. Fructose and sucrose were increased whereas citric acid was decreased during guava fruit ripening.     

Molecular and phenotypic analyses reveal association of diverse Colletotrichum acutatum groups and a low level of C. gloeosporioides with olive anthracnose

Anthracnose (Colletotrichum spp.) is an important disease causing major yield losses and poor oil quality in olives. The objectives were to determine the diversity and distribution pattern of Colletotrichum spp. populations prevalent in olives and their relatedness to anthracnose pathogens in other hosts, assess their pathogenic variability and host preference, and develop diagnostic tools. A total of 128 Colletotrichum spp. isolates representing all olive-growing areas in Portugal and a few isolates from other countries were characterized by molecular and phenotypic assays and compared with reference isolates. Arbitrarily primed PCR data, internal transcribed spacer of rRNA gene and beta-tubulin 2 nucleotide sequences, colony characteristics, and benomyl sensitivity showed Colletotrichum acutatum to be dominant (>97%) with limited occurrence of Colletotrichum gloeosporioides (<3%). Among C. acutatum populations, five molecular groups, A2 to A6, were identified. A2 was widely prevalent (89%), coinciding with a high incidence of anthracnose and environmental conditions suitable to disease spread. A4 was dominant in a particular region, while other C. acutatum groups and C. gloeosporioides were sporadic in their occurrence, mostly related to marginal areas of olive cultivation. C. gloeosporioides, isolated from olive fruits with symptoms indistinguishable from those of C. acutatum, showed same virulence rating as the most virulent C. acutatum isolate from group A2. C. acutatum and C. gloeosporioides isolates tested in infected strawberry fruits and strawberry and lupin plants revealed their cross-infection potential. Diagnostic tools were developed from beta-tubulin 2 sequences to enable rapid and reliable pathogen detection and differentiation of C. acutatum groups.     

A cytochrome P450 gene is differentially expressed in compatible and incompatible interactions between pepper (Capsicum annuum) and the anthracnose fungus, Colletotrichum gloeosporioides

The anthracnose fungus, Colletotrichum gloeosporioides, was previously shown to have an incompatible interaction with ripe-red fruit of pepper (Capsicum annuum). However, the fungus had a compatible interaction with unripe-mature-green fruit. Using mRNA differential display, we isolated and characterized a PepCYP gene expressed in the incompatible interaction. The PepCYP gene encodes a protein homologous to cytochrome P450 proteins containing a heme-binding domain. The expression level of PepCYP is higher in the incompatible interaction than in the compatible interaction, and then remains elevated in the incompatible interaction. In the compatible interaction, the expression of PepCYP is transient. The induction of PepCYP gene is up-regulated by wounding or jasmonic acid treatment during ripening. Analysis of PepCYP expression by in situ hybridization shows that the accumulation of PepCYP mRNA is localized in the epidermal cell layers, but not in the cortical cell layers. An examination of transverse sections of the fruits inoculated with the fungus shows that the fungus invades and colonizes the epidermal cell layers of the unripe fruit at 24 and 72 h after inoculation, respectively, but not those of the ripe fruit. These results suggest that the PepCYP gene product plays a role in the defense mechanism when the fungus invades and colonizes the epidermal cells of fruits in the incompatible interaction during the early fungal infection process.