Molecular characterization and genetic variability studies associated with fruit quality of indigenous mango (Mangifera indica L.) cultivars

Start codon-targeted (SCoT) markers were used for characterization and genetic comparison analysis among 20 mango cultivars (15 indigenous and 5 popular) with respect to fruit quality. Out of 80 SCoT markers used, 19 were able to amplify. These primers produced total 117 loci across 20 cultivars, of which 96 (79.57 %) were polymorphic with an average of 5.05 polymorphic fragments per primer. Out of 19, 17 SCoT primers produced 34 cultivar-specific DNA finger prints. These were 25 unique fragments for identification of 15 indigenous cultivars and 9 fragments for the identification of five popular cultivars. The three SCoT primers—40, 45, and 51 are most informative in identifying mango cultivars as they possess the higher primer index values. The 20 mango cultivars were clustered into two major groups based on the SCoT data analysis with UPGMA. Three indigenous cultivars—Khodi, Amrutiyo, and Kaju and one popular—Dasheri out grouped from other 16 cultivars and shared only minimum similarity (11 %). In clustering pattern, indigenous cultivars—Kaju and Amrutiyo grouped together and shared 37 % similarity with higher boot strapping values (63 %). Clustering pattern is corresponding well with their physical and/or biochemical properties of fruits. The results of principal coordinates analysis (PCoA) analysis were comparable to the cluster analysis. The first three most informative PC components explained 56.61 % of the total variation. In PCoA, three indigenous cultivars—Jamrukhiyo, Chappaniyo, and Sopari appears to be distinct from other 12 indigenous, which be different in fruit size, sugars, ascorbic acids, and carotenoids content. Similarly, popular cultivars—Jamadar and Kesar were also discrete from Alphonso, Dasheri, and Neelum in PCoA. The results demonstrate that the SCoT marker system is useful for cultivar identification and genetic diversity analysis of mango cultivars based on their biological traits.     

几个芒果品种的胚性及多胚苗遗传分析

通过目测,筛选出芒果多胚种子,观察其多胚形态;将多胚种子培育成多胚苗,观察其生长发育状况;取多胚苗和母树叶片进行体细胞染色体数目鉴定及同工酶的分析.结果表明,多胚类群的品种中有单胚现象出现,土芒、吕宋、象牙单胚出现的频率分别为1.04%、3.85%和5.44%;土芒和象牙胚数出现的范围分别为2-6和2-8.多胚种子子叶形态和胚轴的着生位置各异.种子多胚的萌发率、成活率及多胚苗的生长状况均与多胚种子的胚数、子叶发育大小等因素相关.多胚苗体细胞染色体数目是2n=40,未发现染色体的数目变异.POD同工酶分析表明,同一种子的多胚苗间酶谱上存在着差异.     

Development of microsatellite markers for mango (Mangifera indica L.)

Microsatellite markers for mango (Mangifera indica L.) were developed using a genomic library enriched for (GA)_n and (GT)_n dinucleotide repeats. A subset of 41 positive clones was sequenced and primers were designed. Twenty‐eight primer pairs produced polymorphic amplification products for a diversity sample including 15 mango cultivars and two accessions from the related species Mangifera laurina and Mangifera applanata. Nineteen simple sequence repeat (SSR) loci with clear scorable patterns were chosen to study diversity in the mango germplasm bank of Guadalupe (FWI). The number of alleles ranged from three to 13 with observed levels of heterozygosity ranging from 0.059 to 0.857.     

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.     

Biotechnological advances in mango (Mangifera indica L.) and their future implication in crop improvement: a review

Biotechnology can complement conventional breeding and expedite the mango improvement programmes. Studies involving in vitro culture and selection, micropropagation, embryo rescue, genetic transformation, marker-assisted characterization and DNA fingerprinting, etc. are underway at different centers worldwide. In vitro culture and somatic embryogenesis of several different genotypes have been achieved. The nucellus excised from immature fruitlets is the appropriate explant for induction of embryogenic cultures. High frequency somatic embryogenesis has been achieved in some genotypes; however, some abnormalities can occur during somatic embryo germination. Embryo rescue from young and dropped fruitlets can improve the hybridization success in a limited flowering season. Protocols for protoplast culture and regeneration have also been developed. In vitro selections for antibiotic tolerance and fungal toxin resistance have been very promising for germplasm screening. Genetic transformation using Agrobacterium tumefaciens has been reported. Genes that are involved with fruit ripening have been cloned and there have been attempts to deliver these genes into plants. DNA fingerprinting and studies on genetic diversity of mango cultivars and Mangifera species are also being conducted at several research stations. The purpose of this review is to focus upon contemporary information on biotechnological advances made in mango. It also describes some ways of overcoming the problems encountered during in vitro propagation of mango.     

Somatic embryogenesis and plantlet regeneration in mango (Mangifera indica L.)

Summary The ‘Carabao’ or ‘Manila Super’ mango (Mangifera indica L.), a virtually neglected fruit before the advent of KNO₃ flower induction in the early 1970s, is now the third leading Philippine export fruit after banana and pineapple. To apply biotechnology for improvement, a reliable embryogenesis and regeneration protocol is required. We have developed a protocol for somatic embryogenesis and plantlet regeneration in mango: eight strains of ‘Carabao’ and two unidentified varieties, PHL 12384 and PHL 12378. Over 40 batches of nucellar explants from immature fruis (0.75–5.0 cm long) were cultured in vitro from April 1999 to April 2000. Two media were used, MMSE. Mango Medium for Somatic Embryo Induction, Proliferation and Germination and MMPR, Mango Medium for Plantlet Regeneration. These are now routinely used. The protocol is reproducible in 14 other varieties of mango. Shifting the base medium from Gamborg's B5 medium to our own formulation. BP medium (Barba and Pate?a's formulation) effectively controlled browning. Browning has limited the successful in vitro culture of many woody species including the mango. Crop Science Society of the Philippines (CSSP) 2001 Best Paper Award, Asian Agriculture Congress, Westin Philippine Plaza, Manila, Philippines, April 24–27, 2001 and Philippine Fruit Association 2000 Best Poster Award, 8th National Symposium. PCARRD, Los Ba?os, Laguna, Philippines, November 14–16, 2000.     

Changes in peel pigmentation during ripening of mango fruit (Mangifera indica var. Tommy Atkins)

The pigments in the peel of Tommy Atkins mango were analysed at six stages during ripening at 22 ^oC. The loss of green colour and the development of yellow colouration was associated with an almost complete loss of chlorophyll and an increase in carotenoids. Anthocyanin content showed a slight decrease during ripening. An ultrastructural study showed plastids in green fruit with a well developed grana network system. On ripening the chloroplasts underwent extensive disorganisation which was associated with the development of large osmiophilic globules.     

Death and decay in the trees of Mango (Mangifera indica L.)

Death and decay of trees of Mango (Mangifera indica L.) var. Kesar due to fungal infection was studied histologically. Fungal infection in the trees was observed due to various reasons like mechanical injuries in the stem, pruning of the branches, through the inflorescences, attack of Ambrosia beetle and termites. In the initial stage, fungal spores get settled on the flowers due to presence of nectar, followed by their germination and entry of the hypha into peduncle, which gradually spreads into younger branches. The inflorescences were first attacked by Fusarium moniliformis followed by other fungi like Alternaria, Chetomium sp., Aspregillus ellipticus, Aspregillus niger, etc. Fungal mycelia gradually invade the xylem tissues from the top of the branches and spread basipetally ultimately causing death of the infected branches. During monsoon, the crevices on the surface of bark of the healthy plants supported the growth of fungi like Pleurotus, Auricularia, Xyleria, Daldinia sp., and Polyporous sp. The removal of bark from such infected trees revealed minute holes on the surface of the woody cylinder made by Ambrosia beetles. During wet season fungal mycelia makes an easy entry into the xylem through the wounded portion of the stem or pruned branches. Initial entry of the hyphae into xylem was seen through the ray cells. Then the hyphae enter into the lumen of axial elements lining the ray cells through pits and intracellular spaces. The vessel elements located in the xylem (transition zone) between healthy and infected portion were filled with tyloses while axial and ray parenchyma showed heavy accumulation of tannin contents. On the other hand, the infected xylem was also found devoid of reserve metabolites while in normal trees, axial and ray parenchyma showed heavy accumulation of starch grains.     

Structural and rheological properties of polysaccharides from mango (Mangifera indica L.) pulp

The structure and rheological properties of water-soluble polysaccharides from industrialized mango pulp were investigated. Soluble fraction (SF) 2 was heterogeneous on high performance size exclusion chromatography, giving two peaks as determined by multi-angle laser light scattering and refractive index detectors. The presence of starch in SF2 was demonstrated by a positive iodine reaction and by 13C nuclear magnetic resonance (NMR) spectroscopy. The presence of pectic polysaccharides was shown by a calorimetric method, 13C-NMR spectroscopy and carboxyl reduction. The main pectic polysaccharide was polygalacturonic acid; type I rhamnogalacturonan was also detected. Analysis of the rheological properties of SF2 showed a pseudoplastic behavior up to 3 g x l(-1). 'Creep and recovery' tests and analysis performed under a dynamic state revealed a weak gel character for solutions at concentrations of 15, 20 and 30 g x l(-1).     

Plant regeneration from protoplasts of mango (Mangifera indica L.) through somatic embryogenesis

 This report describes a protocol for the regeneration of plants from protoplasts isolated from proembryogenic masses (PEMs) in a suspension culture derived from the nucellar callus of mango (Mangifera indica L. cv 'Amrapali'). The maximum yield (24.6±1.1×10⁶), with 81.04±4.1% viable protoplasts per gram PEMs, was obtained with an enzyme mixture containing 1.2% cellulase, 1.0% hemicellulase and 0.6% pectinase. An optimum density of 5×10⁴ cultured protoplasts per milliliter culture medium was required for the highest frequency (88.89±5.40%) of division. Dividing protoplasts developed into microcalli that proliferated on medium supplemented with growth regulators (auxins or kinetin alone, or auxins with kinetin) and produced somatic embryos after transfer to a growth regulator-free medium. The protocallus on 2,4-D-containing medium produced the maximum number (102.50±6.93) of somatic embryos. Maturation of somatic embryos depended upon the presence, and the nature and combination of growth regulators in the medium during proliferation of the callus. The mature somatic embryos germinated and developed into plants that were transferred to soil. Received: 1 April 1999 / Revision received: 27 July 1999 / Accepted: 23 August 1999     

Nutrient levels in malformed and healthy tissues of mango (Mangifera indica L.)

Samples of malformed and healthy panicles of mango (Mangifera indica L.) as well as leaves and shoots bearing them were collected at different stages of development (fully swollen buds, bud inception, fully grown panicles prior to full bloom and at full bloom) over two consecutive years and were analysed for their macro- and micronutrient status. In addition, malformed and healthy seedlings were collected and analysed. Malformed panicles were found to be significantly higher in N at all the developmental stages except at bud inception. Phosphorus and K also tended to accumulate in malformed panicles at later stages of their development. In general, malformed panicles exhibited lower levels of P, K and Ca than healthy panicles. The differences in levels of Mg and S in malformed and healthy panicles were not significant. All micronutrients were in much lower concentrations in malformed panicles except for Mn which appears to accumulate in malformed panicles particularly at the early stages of development. The leaves on the shoots bearing malformed panicles also showed a tendency to accumulate N, while P, Mg and S were always higher in leaves on shoots bearing healthy panicles. The leaves on shoots bearing healthy panicles had lower levels of Fe, Cu and Mn, whereas levels of Zn and B tended to be higher in leaves on shoots bearing malformed panicles. The nutrient concentration differences between the two kinds of shoots were generally nonsignificant (P=0.05), except for K and S which were significantly lower in shoots bearing malformed panicles. The shoots bearing malformed panicles showed significantly (P=0.05) higher levels of almost all nutrients compared with shoots bearing healthy panicles. Vegetative malformation was found to be associated significantly (p=0.05) with higher amounts of all nutrients except Ca which was significantly higher in healthy seedlings. The present study, therefore, seems to point to lower Ca as one of the pre-disposing factors causing malformation in mango.A part of Ph.D. thesis of the senior author.A part of Ph.D. thesis of the senior author.     

Transcriptome analysis of flowering genes in mango (Mangifera indica L.) in relation to floral malformation

Journal of Plant Biochemistry and Biotechnology - Flowering is a complicated developmental process of physiological and morphological stages under the control of a number of external signals and...     

Seasonal fluctuation in nematode population associated with mango,Mangifera indica L.

Abstract

Mango, Mangifera indica L. Family – Anacardiaceae, an economically important fruit tree, was selected to study the effect of seasonal changes of the population of plant parasitic nematodes viz., Hoplolaimus indicus, Helicotylenchus indicus, Rotylenchulus reniformis, Tylenchorhynchus mashoodi, Tylenchus filiformis and Hemicriconemoides mangiferae around the roots. The population was investigated at 10, 20 and 40 cm depths. It was observed that seasonal fluctuations have a direct effect on the nematode population. The population was larger at 10 cm depth followed by 20 and 40 cm depths. The largest nematode population was observed when the percent soil moisture was high. Both soil temperature and soil moisture were equally important. The soil pH also affected indirectly the nematode population densities.     

Germination and plantlet regeneration from encapsulated somatic embryos of mango (Mangifera indica L.)

 Cotyledonary-stage somatic embryos (3–5 mm in length) originating from nucellar explants of Mangifera indica L. cv. Amrapali were encapsulated individually in 2% alginate gel. The encapsulated somatic embryos (ESEs) germinated successfully on 0.6% agar-gelled medium containing B5 macrosalts (half strength), Murashige and Skoog microsalts (full strength), 3% sucrose and 2.9 μM gibberellic acid. The percentage of germination of ESEs was higher than that of naked somatic embryos of the same size on the same medium. The germinability of ESEs was increased (73.61±7.08%) when the medium was supplemented with full-strength B5 macrosalts. Of the germinating ESEs, 45.83±3.40% developed into plantlets. Abscisic acid at 0.004 and 0.02 μM had no significant influence on germination and plantlet development, but caused a 3-week delay in germination. Well-developed plantlets regenerated from ESEs have been successfully established in soil. Received: 9 February 1998 / Accepted: 22 March 1999     

Ethanolic extract of mango (Mangifera indica L.) malformed inflorescence as effective antifungal agent

Mango (Mangifera indica L.) suffers from floral and vegetative malformation and crop production is seriously affected. The anti-fungal activity of ethanolic extract of malformed mango inflorescence was observed at different concentrations (1000, 2000, 3000, 4000 and 5000 μl/ml) against 10 fungi, viz., Ustilago cynodontis, Cercospora cajani, Sphaerotheca sp., Cercospora sp., Alternaria solani, Bipolaris sp., Helminthosporium sp., Curvularia sp., Fusarium udum and Alternaria cajani. Spore germination of most of the fungi was inhibited at 5000 μg/ml. Some of them were also susceptible at 3000 or 4000 μg/ml concentration. Analysis of phenolic acids by high performance liquid chromatography (HPLC) showed 18 peaks in the extract, but only four could be identified, viz., capachin, gallic, benzoic and cinnamic acids. Because of the high efficacy of ethanolic extract of malformed mango inflorescence, its use under field conditions to control some plant diseases has been suggested.     

Detection of unusual carotenoid esters in fresh mango (Mangifera indica L. cv. 'Kent')

The carotenoid pattern of mango cv. 'Kent' was investigated by LC-(APcI)MS analyses. In solvent extracts from the mesocarp an unusual carotenoid ester was identified as violaxanthin dibutyrate. For unequivocal identification of butyric acid by an independent method, total lipids were isolated by solvent extraction from the fruit flesh and analyzed by GC after saponification and subsequent methylation. Thus, evidence of butyric acid (1.6 area%) was provided. To the best of our knowledge, this is the first report on a xanthophyll dibutyrate in plants. Additionally, further carotenoid peaks were tentatively assigned to 9-cis-violaxanthin and neochrom or luteoxanthin, respectively, by their UV/vis and MS data of the saponified extracts.