Differential gene expression associated with flower development of mango (Mangifera indica L.) varieties with different shelf-life

Mango (Mangifera indica L.) is one of the most important commercial fruit crop grown in many parts of the world. Major challenges affecting mango trade are short shelf-life, high susceptibility to chilling injury, post-harvest diseases and consumer demand for improved fruit quality. The objective of the present study was to reveal the key regulators present in bud and flower tissues during flower development stage, associated with fruit development and affect the shelf-life of the mango fruit. RNA-sequencing of contrasting genotypes having short and long shelf-life, was carried out. Comparative differential expression pathway studies of long shelf-life (Totapuri) and short shelf-life (Bombay Green) mango genotypes revealed a total of 177 highly differentially expressed genes. Out of 177 total genes, 101 genes from endoplasmic reticulum pathway and very few from gibberellins (3) and jasmonic acid (1) pathway were identified. Genes from endoplasmic reticulum pathway like hsp 90, SRC2, DFRA, CHS, BG3 and ASPG1 mainly up regulated in Bombay Green. Uniprotein B9R8D3 also shows up regulation in Bombay Green. Ethylene insensitive pathway gene EIL1 up regulated in Bombay Green. Gene CAD1 from phenylpropanoid pathway mainly up regulated in Bombay Green. A total of 4 SSRs and 227 SNPs were mined from these pathways specific to the shelf-life. Molecular studies of endoplasmic reticulum, phenylpropanoid, ethylene, polygalacturonase and hormone pathways at the time of bud and flower formation revealed key regulators that determine the shelf-life of mango fruit.     

Shrunken-1 encoded sucrose synthase is not required for sucrose synthesis in the maize endosperm

Kernels of wild-type maize (Zea mays L.) shrunken-1 (sh1), deficient in the predominant form of endosperm sucrose synthase and shrunken-2 (sh2), deficient in 95% of the endosperm ADP-glucose pyrophosphorylase were grown in culture on sucrose, glucose, or fructose as the carbon source. Analysis of the endosperm extracts by gas-liquid chromatography revealed that sucrose was present in the endosperms of all genotypes, regardless of carbon supply, indicating that all three genotypes are capable of synthesizing sucrose from reducing sugars. The finding that sucrose was present in sh1 kernels grown on reducing sugars is evidence that shrunken-1 encoded sucrose synthase is not necessary for sucrose synthesis. Shrunken-1 kernels developed to maturity and produced viable seeds on all carbon sources, but unlike wild-type and sh2 kernels grown in vitro, sucrose was not the superior carbon source. This latter result provides further evidence that the role of sucrose synthase in maize endosperm is primarily that of sucrose degradation.     

Diversity analysis,pathogen load and gene expression studies of mango cultivars infected by mango malformation disease

Mango (Mangifera indica L.) is known as “king of fruits” in India. More than 1000 mango varieties are currently cultivated in Indian Sub-continent. However most of the orchards of mango are infected with mango malformation disease (MMD), which every year leads to huge losses in yield of mango in range of 40 to 80?% in India. Till date there is no effective control measure against MMD. Floral Malformation, in contrast to vegetative one, is very virulent and can cause the loss of the entire crop. In the present study, six mango cultivars commonly grown in Gujarat, and all infected with various degrees of MMD were taken for studying their molecular relatedness, pathogen load and defense responsiveness via gene expression to rate whether hybrids or landrace among mango cultivars are better equipped to fight MMD. Genetic diversity analysis was performed using 30 SSR markers in order to bring out clustering pattern among the six cultivars belonging to orchards of Balisana and Prantij, Gujarat. The diversity analysis gave clues to the existence of wide genetic base among the six cultivars. Fungal load studies using Real Time PCR lead to the ranking of cultivars based on maximum and minimum infection load of pathogen. Absolute quantitation studies found that cultivars like Totapuri, Neelam and Amrapali were more resistant to MMD than highly popular cultivars like Kesar. The six mango cultivars were further quantified for pathogen responsiveness with 21 defense responsive genes using Real Time PCR. Among the 21 genes selected for the study, 11 genes were directly part of defense responsive pathways like Phenyl propanoid pathway and jasmonic acid pathway. Gene expression studies aided in ranking mango hybrid like Amrapali having better systemic acquired resistance response as 11 defense responsive genes were found upregulated in this cultivar followed by landrace Neelam which is in fact a parental line of Amrapali. If MMD remains unchecked it may lead to evolution of more virulent strains of Fusarium; propelling devastating consequences in mango cultivation. Hence mango hybrids developed via molecular and expressional screening will fasten process of establishment of resistant mango cultivars.     

Comparative analysis of antioxidant activity,toxicity, and mineral composition of kernel and pomace of apricot (Prunus armeniaca L.) grown in Balochistan,Pakistan

The present study aims to investigate some physical attributes, total phenolics content, total flavonoids content, mineral composition, bioluminescence toxicity assay and antioxidant activity in terms of DPPH, HPS, TAC and FRAP assays in the kernel and pomace samples of six apricot cultivars grown in Balochistan, Pakistan. TFC and TPC determined by the AlCl₃ and Folin-Ciocalteu assays in apricot kernel extracts of six cultivars varied from 1797.5 (Chagali) to 4778.9 (Badoghur) mg QUE/100 g DW and from 1750.0 (Chagali) to 5005.8 (Badoghur) mg GAE/100 g DW. Apricot kernels exhibited higher antioxidant activity than pomace; antioxidant activity in terms of IC₅₀ in kernels ranged from 24.88 to 98.61 μg/ml for DPPH, 334.84 to 516.63 μg/ml for HPS, from 22.02 to 110.80 μg/ml for TAC and from 96.27 to 163.35 μg/ml for FRAP. The apricot kernels showed higher TPC, TFC, bioluminescence toxicity to V. logei and antioxidant activity than the pomace. The correlation analysis demonstrated substantial contributions of polyphenols and flavonoids to antioxidant assays. The sample type was the leading factor affecting the amounts of K, Na, Ca, Fe, and Mn in the tested samples; mineral contents were higher in pomace than kernels. The highest inhibition to V. logei was found in the kernels of Badoghur (IC₅₀ = 1.61 mg/ml). The PCA analysis showed significant contributions of phenolic and flavonoid contents towards antioxidant bioluminescence toxicity assays. Our results suggest Badoghur, Shakarpara and Sardai kernels are rich sources of secondary metabolites and possess remarkable antioxidant and antiluminescence activity and can make a significant contribution to the treatment and prevention of chronic health problems.     

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.     

Life history traits of Blaptostethus pallescens (Hemiptera: Anthocoridae), a candidate for use in augmentative biological control in Egypt

Blaptostethus pallescens Poppius (Hemiptera: Anthocoridae) is an abundant native predator in mango orchards and other cropping systems in Egypt. To determine suitable mass-rearing conditions for this little-studied species, we assessed some of its biological characteristics. Testing its thermal response at three constant temperatures (20, 25, 30 °C), showed that immature development time and adult longevity decreased with increasing temperature. Reproductive success of individual females was greatest when reared at 25 °C (84.3 ± 3.1 eggs) rather than at 20 °C (46.6 ± 2.0 eggs) or 30 °C (65.2 ± 2.5 eggs). Although B. pallescens reared at 25 °C had a significantly higher net reproductive rate (R ₀), which may be attributed to their relatively rapid development and high fecundity, we argue that 30 °C seems to be more convenient for rearing B. pallescens, as mean generation time (T) and doubling time (DT) are clearly shorter, thus more individuals could be reared per unit of time at 30 °C. Mating significantly reduced male and female longevity, as unmated adults lived 25–45 % longer than mated individuals did. Unmated females did not lay eggs, suggesting that mating is a prerequisite for egg maturation. Adult males and females performed best, in terms of longevity, when fed Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs instead of non-prey diets. However, diets of plant sap or pollen could sustain adults in times of limited egg availability. Because its biology is similar to that of other subtropical anthocorids already reared for augmentative releases, B. pallescens may be amenable to mass-rearing using already established techniques. Therefore, B. pallescens could be used to improve augmentative biological control in crops such as mango or maize in Egypt where it already naturally occurs, and therefore would not engender concerns over non-target effects that an exotic, generalist biological control agent would.     

Conversion of mango kernel starch to glucose syrups by enzymatic hydrolysis

In the present investigation, the possibility of utilizing the starch present in mango seed (which are thrown away as waste) kernels for the production of glucose syrups by enzyme-enzyme hydrolysis has been studied. Under the conditions of operation, particles less than 90 microns in size showed maximum conversion at i) -amylase concentration=0.06% (v/v), pH=6.5 and temperature 95°C and ii) glucoamylase concentration=0.8% (v/v), pH=4.5 and temperature=60°C.     

Volatile flavour components of mango fruit

An essence of fresh Venezuelan mango fruit obtained by well-established procedures possessed the characteristic aroma of the fruit. It was analysed by GC/MS using both EI and Cl. The fruit produced a relatively small quantity of aroma volatiles (ca 60 μg/kg fresh fruit), less than that obtained from many similar tropical fruits. Terpene hydrocarbons comprised ca 68% of the sample, eight monoterpenes contributing ca 54% and four sesquiterpenes contributing ca 14%. Important constituents included α-pinene, car-3-ene, limonene, γ-terpinene, α-humulene, β-selinene, acetophenone, benzaldehyde and a dimethylstyrene. Car-3-ene (26%) was the major constituent, and on odour evaluation of separated components at an odour port during GC, the peak due to this compound was described as having an aroma of mango leaves. This compound has not previously been detected among mango volatiles. The only other component providing mango aroma was a dimethylstyrene, and this too is a new mango volatile.     

Regulation of Embryo Dormancy by Manipulation of Abscisic Acid in Kernels and Associated Cob Tissue of Zea mays L. Cultured in Vitro

Sectors of Zea mays cobs, with and without kernels were cultured in vitro in the presence and absence of fluridone. Cultured kernels, cob tissue, and embryos developed similarly to those grown in the field. Abscisic acid (ABA) levels in the embryos were evaluated by enzyme-linked immunosorbant assay. ABA levels in intact embryos cultured in the presence of fluridone were extremely low and indicate an inhibition of ABA synthesis. ABA levels in isolated cob tissue indicate that ABA can be produced by cob tissue. Sections containing kernels cultured in the presence of fluridone were transferred to medium containing fluridone and ABA. Dormancy was induced in more than 50% of the kernels transferred from 13 to 15 days after pollination, but all of the kernels transferred at 16 days after pollination or later were viviparous. ABA recovered from kernels that were placed in medium containing fluridone and ABA suggest that ABA can be transported through the cob tissue into developing embryos and that ABA is required for induction of dormancy in intact embryos.     

Peanut Pod Invasion by Aspergillus flavus in the Presence of Meloidogyne hapla

''Argentine'', ''Early Runner'' and ''Florigiant'' peanut cultivars were grown in methyl bromide treated soil in field microplots inoculated with: (i) Aspergillus flavus or (ii) A. flavus + Meloidogyne hapla. Nematode infection produced heavy root galling and light pod galling equally on all cultivars. A. flavus, A. niger, Cephalosporium spp., Colletotrichum sp., Curvularia spp., Fusarium spp., Penicillium spp. and Trichoderma viride were isolated from shells and kernels. A significantly greater incidence and density of A. flavus was obtained from kernels of plants inoculated with both organisms than from kernels of plants receiving only the fungus. Differences were not significant, however, for incidence and density of A. flavus in shells or for the total of all fungal propagules in shells and kernels. Shells of ''Early Runner'' contained significantly greater incidence and density of A. flavus than the other two cultivars; also, kernels of this cultivar contained more fungal propagules than kernels of ''Argentine.'' A significantly larger number of total fungi was isolated from kernels of ''Argentine'' than from ''Florigiant.'' Aflatoxins were found only in two shell samples and not in kernels.     

Distribution of Abscisic Acid in Maize Kernel during Grain Filling

The distribution of abscisic acid (ABA) within maize (Zea mays L.) kernels was studied in kernels from nontreated plants, from plants in which assimilate supply had been altered by source/sink manipulations, and in kernels cultured in vitro on ABA-free media. Prior to growth of the embryo, both the pedicel/placento-chalazal complex and the endosperm contained high concentration of ABA; however, the quantity of ABA in these tissues declined as the concentration in the embryo increased during the early stages of embryo growth. Peaks in the levels of ABA appeared to occur prior to and not concurrent with physiological events during grain filling. During most of the grain filling period, ABA concentration in the embryo was higher than that found in other kernel components. Altering assimilate supply by partial defoliation at two stages of development resulted in variable and transient effects on the relative distribution and concentration of ABA in kernel components. The concentration and distribution of ABA among components of kernels grown in vitro was similar to that observed for field-grown kernels. On the basis of these findings, in situ synthesis of ABA by kernel components is implicated and the putative role of ABA in the regulation of kernel development is discussed.     

Growth and composition of maize kernels cultured in vitro with varying supplies of carbon and nitrogen

This study employed in vitro seed culture to determine how C and N supply influence the growth (i.e. starch accumulation) and protein composition of maize (Zea mays L.) endosperm. Immature kernels were grown to maturity on liquid medium containing various concentrations of C (sucrose at 234 millimolar [low] and 468 millimolar [high]) and N (amino acid mixture ranging in N from 0 to 144 millimolar). Low C supply limited starch, but not N, accumulation in the endosperm. With high C, endosperm starch and protein content increased concomitantly as N supply increased from 0 to 13.4 millimolar. Endosperm growth was unaffected by additional N until concentrations exceeding approximately 72 millimolar reduced starch accumulation. A similar inhibition of starch deposition occurred with lower N concentrations when kernels were grown with low C. Endosperm total N content reached a point of saturation with approximately 36 millimolar N in the medium, regardless of C supply. Zein synthesis in the endosperm responded positively across all N levels, while glutelin content remained static and albumin/globulin proteins were reduced in amount when N supply was greater than 36 millimolar. A reciprocal, inverse relationship was observed in mature endosperm tissue between the concentrations of free amino acids and soluble sugars. Our data suggest that under N stress starch and protein accumulation in the endosperm are interdependent, at least in appearance, but are independent otherwise.     

Tropical and subtropical fruits in florida (other than citrus)

Avocado and mango are the leading commercial fruits of South Florida. Lychee, guava, pineapple, bananas, papayas, and persimmon, are of minor economic value, while sapodilla, macadamia nut, Barbados cherry and the Mysore black raspberry show promise of commercial development. All of these fruits and many more are grown as dooryard plants. The fact that such a large number of tropical and subtropical fruits can be grown in South Florida has made the subject of tropical pomology a very important one for this area.     

Notes on distribution,propagation, and products ofBorassus Palms (Arecaceae)

The palmyra, or toddy, palm (Borassus flabellifer L., Arecaceae) grows wild from the Persian Gulf to the Cambodian-Vietnamese border; is commonly cultivated in India, Southeast Asia, Malaysia and occasionally in other warm regions including Hawaii and southern Florida, where there are also a number of specimens of the similarB. aethiopum Mart. In addition to the sweet sap from the inflorescence and the many products of the leaves, trunk and underground seedlings, a thin orange pulp coating the fibers of the mature fruit is consumed fresh or dried as a paste. The large seeds, when immature, before the shell hardens, contain jelly-like kernels esteemed for food. These kernels, whole or sliced and canned in thin, clear sirup, are being exported from Thailand and are appearing in Asiatic markets across the United States. Mature kernels are hard as ivory. A number ofBorassus palms have succumbed to lethal-yellowing disease in Florida.     

Mango - Postharvest Biology and Biotechnology

Mango is one of the choicest fruits in the world and popular due to its delicate taste, pleasant aroma and nutritional value. Mango is indigenous to north-east India and north Burma, but now grown in over 90 countries. In the past two decades, mango production has increased appreciably with international trade jumping approximately four-fold valued close to US$ 950 million. Mango belongs to the category of climacteric fruits and its ripening is initiated and proceeded by a burst in ethylene production and a dramatic rise in the rate of respiration. Although there are a few hundred mango cultivars grown in the Indian subcontinent and other parts of the world, the most popular cultivars are generally highly perishable and ripen within 7 to 9 days of harvest at ambient temperature. Currently, the export potential and international trade of mango is limited due to several factors such as its perishable nature, disease and pest infestation, and susceptibility of certain premium cultivars to chilling injury when stored at low temperatures. Efforts are ongoing to develop technologies for improved storage and packaging, and overcome limitations encountered during storage and transit. Controlled atmosphere (CA) and hypobaric storage of mango are powerful means to overcome its perishable nature. The composition of CA varies among cultivars to ensure its original taste, flavor and aroma. Edible coating on the fruit skin may further cut down the rate of deterioration. Recently, significant advances have been made in understanding ripening characteristics of mango at the molecular level. Candidate genes related to ethylene biosynthesis and signalling, cell wall modification, aroma production and stress response have been cloned and characterized for future use in mango improvement. Efforts are also being made to establish a suitable transformation and plant regeneration system so that transgenic mango with added value and increased shelf life for long distance transportation could be developed.     

Responses of embryogenic mango cultures and seedling bioassays to a partially purified phytotoxin produced by a mango leaf isolate of Colletotrichum gloeosporioides penz

Summary Colletotrichum gloeosporioides Penz., the causal agent of mango anthracnose, produces a phytotoxin in vitro. The partially purified phytotoxin, presumably colletotrichin, caused anthracnose-like symptoms on young mango leaves, was toxic to embryogenic suspension cultures of two mango cultivars, ‘Hindi’ and ‘Carabao,’ and inhibited in vitro seed germination of two nonhosts, lettuce and tobacco. There were linear relationships between concentration of the partially purified phytotoxin and mortality of mango embryogenic cultures. Embryogenic cultures grown in the presence of the partially purified phytotoxin showed significantly lower growth rates than the controls. Similarly, embryogenic cultures grown in the presence of 40% (vol/vol) fungal culture filtrate showed significantly lower growth rates than unchallenged controls. Medium containing 40% (vol/vol) Czapek-Dox fungal broth did not reduce growth of embryogenic cultures, indicating the production of phytotoxin in vitro. The results suggest that either fungal culture filtrate or purified phytotoxin can be used as in vitro selection agents to screen for resistance to this fungus.     

Comparison of the volatile components of some mango cultivars

Three cultivars of mango from Sri Lanka (Jaffna, Willard and Parrot) were analysed for their volatile aroma components. The total concentrations of volatiles obtained were ca 251, 422 and 628 μg per kg of fresh fruit, respectively. Terpenes were the main volatiles of all three cultivars, with monoterpene hydrocarbons contributing 50–63 % w/w of the total volatiles and sesquiterpene hydrocarbons 14–19 %. Whilst the major volatile of Jaffna mango was cis-β-ocimene (38 %), α-terpinolene was the major volatile of the other two cultivars (32 % and 35 %). Esters were produced by all cultivars (2–16 %), Jaffna yielding most, the majority being unsaturated (12 %). Willard mango gave particularly high levels of non-terpene hydrocarbons (19 %), including a range of six long-chain alkanes (8 %), not detected in the other cultivars.     

Regulation of Fumonisin B1 Biosynthesis and Conidiation in Fusarium verticillioides by a Cyclin-Like (C-Type) Gene, FCC1

Fumonisins are a group of mycotoxins produced in corn kernels by the plant-pathogenic fungus Fusarium verticillioides. A mutant of the fungus, FT536, carrying a disrupted gene named FCC1 (for Fusarium cyclin C1) resulting in altered fumonisin B₁ biosynthesis was generated. FCC1 contains an open reading frame of 1,018 bp, with one intron, and encodes a putative 319-amino-acid polypeptide. This protein is similar to UME3 (also called SRB11 or SSN8), a cyclin C of Saccharomyces cerevisiae, and contains three conserved motifs: a cyclin box, a PEST-rich region, and a destruction box. Also similar to the case for C-type cyclins, FCC1 was constitutively expressed during growth. When strain FT536 was grown on corn kernels or on defined minimal medium at pH 6, conidiation was reduced and FUM5, the polyketide synthase gene involved in fumonisin B₁ biosynthesis, was not expressed. However, when the mutant was grown on a defined minimal medium at pH 3, conidiation was restored, and the blocks in expression of FUM5 and fumonisin B₁ production were suppressed. Our data suggest that FCC1 plays an important role in signal transduction regulating secondary metabolism (fumonisin biosynthesis) and fungal development (conidiation) in F. verticillioides.     

Relationship between Meloidogyne arenaria and Aflatoxin Contamination in Peanut

Damaged and developing kernels of peanut (Arachis hypogaea) are susceptible to colonization by fungi in the Aspergillus flavus group which, under certain conditions, produces aflatoxins prior to harvest. Our objective was to determine whether infection of peanut roots and pods by Meloidogyne arenaria increases aflatoxin contamination of the kernels when peanut is subjected to drought stress. The experiment was a completely randomized 2-x-2 factorial with 6 replicates/treatment. The treatment factors were nematodes (plus and minus M. arenaria) and fungus (plus and minus A. flavus inoculum). The experiment was conducted in 2001 and 2002 in microplots under an automatic rain-out shelter. In treatments where A. flavus inoculum was added, aflatoxin concentrations were high (> 1,000 ppb) and not affected by nematode infection; in treatments without added fungal inoculum, aflatoxin concentrations were greater (P ≤ 0.05) in kernels from nematode-infected plants (1,190 ppb) than in kernels from uninfected plants (79 ppb). There was also an increase in aflatoxin contamination of kernels with increasing pod galling (r² = 0.83 in 2001, r² = 0.43 in 2002; P ≤ 0.04). Colonization of kernels by A. flavus increased with increasing pod galling (r² = 0.18; P = 0.04) in 2001 but not in 2002. Root-knot nematodes may have a greater role in enhancing aflatoxin contamination of peanut when conditions are not optimal for growth and aflatoxin production by fungi in the A. flavus group.