Cyanide produced with ethylene by ACS and its incomplete detoxification by ß-CAS in mango inflorescence leads to malformation.

Malformation of mango inflorescences (MMI) disease causes severe economic losses worldwide. Present research investigates the underlying causes of MMI. Results revealed significantly higher levels of cyanide, a by-product of ethylene biosynthesis, in malformed inflorescences (MI) of mango cultivars. There was a significant rise in ACS transcripts, ACS enzyme activity and cyanide and ethylene levels in MI as compared to healthy inflorescences (HI). Significant differences in levels of methionine, phosphate, S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, ascorbate and glutathione, and activities of dehydroascorbate reductase and glutathione reductase were seen in MI over HI. Further, a lower expression of ß-cyanoalanine synthase (ß-CAS) transcript was associated with decreased cellular ß-CAS activity in MI, indicating accumulation of unmetabolized cyanide. TEM studies showed increased gum-resinosis and necrotic cell organelles, which might be attributed to unmetabolized cyanide. In field trials, increased malformed-necrotic-inflorescence (MNI) by spraying ethrel and decreased MNI by treating with ethylene inhibitors (silver and cobalt ions) further confirmed the involvement of cyanide in MMI. Implying a role for cyanide in MMI at the physiological and molecular level, this study will contribute to better understanding of the etiology of mango inflorescence malformation, and also help manipulate mango varieties genetically for resistance to malformation.

Role of ethrel in causation of floral malformation in mango cv. Amrapali: a scanning electron microscopy study.

Floral malformation is a main constraint to reduce fruit yield in mango plants. Recently, we report on the role of putrescine in normalizing the functional morphology of mango flower by reducing various adverse effects of ethylene. Here, ethrel, an ethylene releasing compound, was exogenously applied to mango plant cv Amrapali to evaluate the response of flower development under high level of ethylene. Scanning electron microscopy (SEM) study showed that ethrel treated flowers were observed to progressively be deformed and remain unbloom. The flower buds were not distinguishable and flower parts such as petals, sepals, anther and stigma were not properly developed. The stamen showed fused anther lobes and carpel depicted curved style with pointed stigma. The findings of present study suggest the involvement of ethylene to abort the functional morphology of flower and thereby development of malformation.

Fusarium mangiferae associated with mango malformation in the tarai region of the Uttarakhand state of India.

Mango malformation is the most dangerous disease to mango worldwide. There are hints that Fusarium mangiferae might be one of the probable casual agents of disease. Recently, we reported on Fusarium isolates obtained from the mango tarai region of Uttarakhand acquiring morphological features of F. mangiferae. Here, further confirmation of Fusarium isolates were made by PCR amplification using primers specific to the translation elongation factors 1α and ß-tubulin gene of F. mangiferae. Further, SDS-PAGE and RAPD profiles showed genetic variability among isolates of F. mangiferae. This study provides further direct evidence of involvement of different strains of F. mangiferae in malformation diseases of mango in the tarai region of the Uttarakhand state.

Low temperature stress ethylene and not Fusarium, might be responsible for mango malformation.

Malformation is arguably the most crucial disease of mango (Mangifera indica L.). The etiology of the disease has not yet been successfully resolved. Here, we quantified the endogenous ethylene content in malformed and healthy vegetative and floral tissues of mango cultivars viz., Amrapali, Bombay green, Chausa, Dushehri and Mallika. Levels of ethylene were higher in malformed vegetative and floral tissues as compared with that of healthy tissues at both prior to full bloom and full bloom stages. The study also revealed that isolates of Fusarium dissected from mango exhibited most morphological similarities to the accepted standard features of Fusarium mangiferae. The growth dynamic of F. mangiferae were evaluated with varying temperatures ranging from 5 to 40 °C. Temperatures of 25 °C, 30 °C and 35 °C were better suited for growth of F. mangiferae than temperatures of 20 °C or 40 °C. Conidium germination of F. mangiferae was maximum at 30 °C and minimum at <15 °C. World-wide occurrence of mango malformation showed its most severity at 10-15 °C temperature range. Stress ethylene level is higher in diseased tissue at the same temperature range where growth of Fusaria is found to be completely restricted. The present study provides direct evidence that low temperature induced 'stress ethylene' is potentially responsible for the disease while on the other hand Fusarium role in the disease either through toxic principle or malformation inducing principle is not conclusive at <15 °C and is rather out of question.

First evidence of ethylene production by Fusarium mangiferae associated with mango malformation.

Malformation is arguably the most crucial disease of mango (Mangifera indica L.) at present. It is receiving great attention not only because of its widespread and destructive nature but also because of its etiology and control is not absolutely understood. Recently, Fusarium mangiferae is found to be associated with mango malformation disease. There are indications that stress ethylene production could be involved in the disease. Here we have shown the first direct evidence of production of ethylene in pure culture of F. mangiferae obtained from mango. The study also revealed that all the isolates dissected from mango acquire morphological features of F. mangiferae showing most similarity to the features of species with accepted standard features. The isolates of F. mangiferae from mango were observed to produce ethylene in significant amounts, ranging from 9.28-13.66 n mol/g dry wt/day. The findings presented here suggest that F. mangiferae could contribute to the malformation of mango by producing ethylene and probably stimulating stress ethylene production in malformed tissue of mango. Ethylene might be produced through 2-oxoglutarate-dependent oxygenase-type ethylene-forming-enzyme (EFE) pathway in Fusarium sp, which needs to be investigated.

In vitro: Response of plant growth regulators and antimalformins on conidia germination of Fusarium mangiferae and incidence of mango malformation.

Mango malformation is the most important and threatening disease of recent times, primarily because of persistent lacuna in complete understanding of its nature. Diverse Fusarium spp, including F. mangiferae, were found to be associated with the disease. Here, F. mangiferae from mango cv Dashehri was morphologically characterized. Typically, oval-shaped microconidia without septum and crescent-shaped macroconidia with 3-septate were more often observed, whereas not a single chlamydospore was detected. The length and width of micro- and macro-conidia were 7.5, 55, 3.2, and 3.5, respectively. The plant growth regulators such as NAA, GA3, BAP and ethrel were found to induce in vitro germination of conidia of F. mangiferae after 12 h. In contrast, antimalformin silver nitrate (AgNO3) inhibits conidial germination in vitro and none of conidia was germinated beyond 500 ppm, however antimalformin glutathione was highly effective in stimulating conidial germination of F. mangiferae in vitro at > 1000 ppm after 24 h. We observed that the response of F. mangiferae to germinate the conidia in vitro under influence of plant growth regulators and antimalformins is not coincided with earlier findings of reduced disease incidence by exogenous application of these compounds. The present findings do not authenticate the involvement of F. mangiferae in the disease, however hormonal imbalance, most probably ethylene, might be responsible for deformed functional morphology of panicle. Further, a signal transduction mechanism of stress-stimulated ethylene imbalance causing physio-morphological changes in reproductive organs of mango flower and thereby failure of fertilization and fruit set, which needs to be investigated.