Melon Genotypes Research Articles

Estimation of phenotypic divergence in a collection of muskmelon germplasm, including Indian and exotic populations

Seventy melon genotypes, comprising of landraces, inbred lines and cultivars collected from diverse geographical locations, were evaluated for 17 morphological traits for determining the patterns of genetic diversity and relationship among genotypes. The genetic diversity based on quantitative and qualitative characters of genotypes was assessed by D2 analysis. The Euclidean’s distance-based clustering divided the germplasm in to nine clusters. Though the analysis of 17 descriptors distinctly separated momordica and wild accessions from cantalupensis, reticulatus, inodorus accessions, but it was not able to differentiate cantalupensis, reticulatus, inodorus group accessions. The inter-cluster distance varied from 759.6 between clusters 1 and 2 to 5194.5 between clusters 4 and 8. The maximum intra-cluster distance matrix was observed in cluster 9 (728.5), while cluster 6 exhibited the minimum intracluster variation (344.0). Maximum number of genotypes was grouped in cluster 1 and 7 i.e. 15 in each group. The genotype “Canary Yellow-2” present in cluster 4 did not grouped in any cluster. Overall, the clustering patterns exhibited great diversity in the germplasm for certain traits(fruit morphology and quality characteristics) which can be utilized for broadening the genetic base of muskmelon germplasm and for developing specialty melon cultivars/ hybrids with unique traits such as high beta carotene, sweetness and extended shelf life.

Drought stress mitigation with humic acid in two Cucumis melo L. genotypes differ in their drought tolerance

ABSTRACT Different responses of two melon (Cucumis melo L.) genotypes (Semame, drought and salt-tolerant and Ananas, drought and salt-sensitive) to drought stress with or without humic acid (HA) treatment were studied. The experiment was carried out under greenhouse conditions. The experimental design was two factorial randomized block with 4 replicates. HA treatment increased the shoot fresh and dry weights and leaf area of both genotypes under drought stress. HA stimulated accumulation of K and Ca ions, chlorophyll (SPAD value) and antioxidant enzyme activity (superoxide dismutase-SOD, catalase-CAT and glutathione reductase-GR) in both genotypes. This effect was more clear in the Semame genotype than in Ananas.As a result, HA treatment has been proved to influence the ability of melon genotypes to cope with drought stress and to increase their tolerance.

Fruit quality assessment and characterization of melon genotypes

ABSTRACT Local melon germplasm are sources of natural variation with great potential, unfortunately, they are being replaced by hybrids and modern high-yielding disease-resistant/tolerant varieties. To recognize, and encourage re-use, local melon genotypes previously selected for resistance to Fusarium wilt disease, were evaluated for morphological, nutritional and biochemical traits. The evaluated melons expressed a wide range of phenotypes for morphological fruit characteristics, taste, and composition. Differences occurred between botanical groups and within the same group. The “Galaoui” genotype of reticulatus botanical group presented the highest β-carotene value (6.6 mg∙kg−1 FW) and the highest flavonoid content (45.45 mg CE∙kg−1 FW). The “Dziri” genotype of inodorus botanical group had the highest phenolic content (896.67 mg GAE∙kg−1 FW); genotype Rupa of cantalupensis group had the highest antioxidant activity value (14.61%). Genotype Asli of inodorus group had the highest vitamin C content (0.15 g/100mL fresh juice). These genotypes might be appropriate for breeding program to increase the functional properties of commercial varieties. Moreover, principal component analysis showed a great dispersion of melon genotypes. The hierarchical ascending classification indicated genotypes were sorted into four cluster groups based on fresh weight, fruit diameter, total soluble solids, total phenolic content, and β-carotene contents. The diversity identified in this study, especially from the four group types, showed that some Tunisian genotypes are featured with several morphological and biochemical characteristics, they had melon quality exceeding commercial hybrids, and could be further exploited as a promising material for the creation of new improved varieties.

Molecular characterization of Acidovorax citrulli strain NIHHS15-280 causing bacterial fruit blotch disease in Korea and screening of resistance sources in melon

Bacterial fruit blotch (BFB), caused by Acidovorax citrulli, is an economically devastating disease in melon. Molecular characterization of causal strains and identification of sources of resistance are necessary to control this important disease. Many strains of BFB were identified by partial sequencing and documented in GenBank. Recently, there was an outbreak of BFB strain NIHHS15-280 in melon fields (Goksung, Jeonnam, Korea) that severely hampered melon production in Korea. For this reason, we characterized NIHHS15-280 by partially sequencing the 16S rRNA gene using PCR-based markers. The partial sequence of the 16S rRNA gene of NIHHS15-280 was much more similar to A. citrulli strains associated with melon and watermelon hosts than other cucurbit hosts. A PCR-based assay identified the strain (NIHHS15-280) as belonging to A. citrulli group I, which predominantly infects non-watermelon cucurbits (virulent to melon). Thirty-five melon genotypes were screened against NIHHS15-280 in a detached-leaf inoculation assay. The lack of visible disease symptoms 11 days after inoculation in lines PI353814 and PI140471 indicated these to be highly resistant lines, whereas moderate resistance was observed in lines PI420145 and SCNU1154 and cultivars ME6 and ME5. Also, the most resistant lines were corroborated by a fruit inoculation assay. The characterized A. citrulli strain NIHHS15-280 (group I) and identified resistant lines will serve as a valuable resource for further genetic studies and improvement of BFB resistance in melon.

In vitro callus induction and shoot regeneration potentials in some snake melon accessions collected from different regions in Egypt

Several snake melon genotypes are grown in different locations in Egypt. However, the relationships and the degree of relatness among these genotypes are not well documented. This study was carried out with the aim to classify different Egyptian snake melon genotypes based on the in vitro callus induction and shoot regeneration potential. Nine snake melon accessions (acc.) were collected from different regions in Egypt, including acc.3 (Damietta), acc.7 (Bany Swief), acc.8 (Fayoum), acc.9 (Giza), acc.11 (Menia), acc.14 (Sohag), acc. 15 (Behaira-Wady Elnatron), acc. 17 (Ismailia) and acc. 18 (Behaira-Badr Center). Twenty seeds from each accession were sterilized and cultured in vitro on MS medium basal salt and vitamins for 4 weeks. Segments (4x4mm each) from cotyledon were used as explants and cultured on MS medium amended with 2.0 mg/L TDZ for callus induction. The formed callus was sub cultured onto MS medium amended with 3.0 mg/L BA+4ml Hyaluronic acid for shoot regeneration. Results indicated that the degree of callus formation was different among the different accessions. Based on callus growth potential measured as fresh weight, snake melon accessions could be ranked as: acc.9> acc.18> acc.3= acc.11> acc.17> acc.15> acc.8= acc.14> acc.7. Significant differences among the accessions were also observed for their shoot regeneration potential from callus. The highest number of shoots per explant was recorded in acc. 18 (ave.12.6 shoots), followed by acc.9, 11, and 7 which produced an average of 10.8, 10.4, and 9.8 shoots, respectively. Results suggested that snake melon genotypes with high callus induction had also high regeneration capacity. In addition, these accessions could have different genetic background, which might help in future breeding programs to improve plant and agronomic traits. The current in vitro callus induction and shoot regeneration technique in snake melon will also aid in future effort for germplasm preservation of accessions with unique characteristics.

Yield Stability and Resistance of Five Melon Genotypes (Cucumis melo L.) Cultivated On Three Altitude Areas

The study was conducted to determine the yield stabilities of five melon varieties tested at three altitude areas and to obtain a variety of melon whose yield and stability exceed at three altitude areas. The study examined five melon varieties, namely 'Golden Aroma', 'Ladika', King Melon, Bravo, and M19 at three altitudes, namely Jombang (lowland 52 m dpl), Jiput (mediumland 460 m dpl), and Bojong (highland 776 m dpl) using a complete randomized block design (RAKL) with four replicates (blocks). The study was conducted during the rainy season in October 2016-January 2017 (in Jombang), November 2016-February 2017 (in Jiput), and March 2017-June 2017 (in Bojong). The results showed that the varieties that had yield per hectare and genotype resistance to the environment exceeded the general average were the varieties of King Melon and Bravo, although both of them were unstable at three altitude areas. Average yield per hectare for King Melon (19.33 ton/ha) and Bravo (18.21 ton/ha) and genotype resistance to the environment for King Melon (65.42%) and Bravo (56.25%) were superior to the commercial varieties of 'Golden Aroma' and 'Ladika' and able to compete with national melon yields in 2013 (17.71 ton/ha) and 2014 (18.37 ton/ha).

Texture diversity in melon (Cucumis melo L.): Sensory and physical assessments

Melon (Cucumis melo L.) is a commercially important horticultural crop worldwide that exhibits extensive phenotypic and genetic variation. Texture is one of the key attributes defining melon fruit quality and overall consumer preference. The aim of this research was two-fold: first, to characterize and compare differences in fruit sensory and instrumental textural properties among a diverse panel of melon fruit genotypes (to determine if each methodology is capable of discriminating among genotypes independently); and secondly, to assess the correlations between texture-related sensory attributes and instrumental parameters. During two production seasons, fruit from 10 melon genotypes with diverse textural characteristics were harvested at optimal commercial maturity and stored for six days at 5 °C plus one day at room temperature, then analyzed for textural instrumental and sensory properties. Both methodologies detected significant and reproducible differences in texture among the assayed melon genotypes. Furthermore, texture-related sensory attributes of firmness, crunchiness, and juiciness significantly correlated with several parameters obtained through the instrumental assessment of texture by using different probes of the texture analyzer instrument, independently. Our results indicate that texture-related attributes and overall fruit quality improvement could be accelerated significantly by the application of instrumental measurements to select for phenotypes that highly associate with consumer perception, decreasing costs and time investments.

CmVPS41 Is a General Gatekeeper for Resistance to Cucumber Mosaic Virus Phloem Entry in Melon.

Melon production is often compromised by viral diseases, which cannot be treated with chemicals. Therefore, the use of genetic resistances is the main strategy for generating crops resistant to viruses. Resistance to Cucumber mosaic virus (CMV) in melon is scarcely described in few accessions. Until recently, the only known resistant accessions were Freeman’s Cucumber and PI 161375, cultivar Songwhan Charmi (SC). Resistance to CMV in melon is recessive and generally oligogenic and quantitative. However, in SC, the resistance to CMV strains of subgroup II is monogenic, depending only on one gene, cmv1, which is able to stop CMV movement by restricting the virus to the bundle sheath cells and preventing a systemic infection. This restriction depends on the viral movement protein (MP). Chimeric viruses carrying the MP of subgroup II strains, like the strain LS (CMV-LS), are restricted in the bundle sheath cells, whereas those carrying MP from subgroup I, like the strain FNY (CMV-FNY), are able to overcome this restriction. cmv1 encodes a vacuolar protein sorting 41 (CmVPS41), a protein involved in the transport of cargo proteins from the Golgi to the vacuole through late endosomes. We have analyzed the variability of the gene CmVPS41 in a set of 52 melon accessions belonging to 15 melon groups, both from the spp melo and the spp agrestis. We have identified 16 different haplotypes, encoding 12 different CmVPS41 protein variants. Challenging members of all haplotypes with CMV-LS, we have identified nine new resistant accessions. The resistance correlates with the presence of two mutations, either L348R, previously found in the accession SC and present in other three melon genotypes, or G85E, present in Freeman’s Cucumber and found also in four additional melon genotypes. Moreover, the new resistant accessions belong to three different melon horticultural groups, Conomon, Makuwa, and Dudaim. In the new resistant accessions, the virus was able to replicate and move cell to cell, but was not able to reach the phloem. Therefore, resistance to phloem entry seems to be a general strategy in melon controlled by CmVPS41. Finally, the newly reported resistant accessions broaden the possibilities for the use of genetic resistances in new melon breeding strategies.

Application of water-saving treatments reveals different adaptation strategies in three Iranian melon genotypes

Melon is a major horticultural crop in Iran, where it is grown in semi-arid areas in irrigated fields. Concerns about future water security stimulate the development of water-saving irrigation practices, which requires a better understanding of melon physiology under reduced water supply. To this end, one variety of muskmelon, “Samsoury”, and two varieties of the Dudaim group, “Birjand” and “Kerman”, were grown in pots in a greenhouse with three water management treatments, consisting in refilling soil water to field capacity at different intervals: three days (T3), six days (T6) and nine days (T9). Leaf water status declined when longer refilling intervals were applied, but all genotypes maintained the same leaf turgor pressure irrespective of the treatment. Turgor was maintained both through osmotic adjustment (OA) and change in cell wall elasticity. Samsoury showed the highest OA in T3 and T6, while variation in cell wall elasticity was more evident in Birjand and Kerman. Samsoury showed also the highest photosynthesis rate (An) and stomatal conductance (gs) in T3 and T6, but the three genotypes exhibited similar values in T9. In the Dudaim-types An and gs declined linearly as soon as water potential started to decrease, while Samsoury was able to keep gas-exchange parameters stable down to a water potential value of –11.5 bars, to steeply decrease thereafter. Finally, Samsoury had the least reduction in biomass allocated to epigeic part, and allocated a higher proportion of biomass to roots in response to water restriction. From all the results, it is concluded that Samsoury performs better when water-saving treatments allow low to moderate water stress, thanks to its anisohydric-like behaviour, consisting in a looser coupling of stomatal closure with water potential, which ensures higher carbon accumulation under fluctuating water supply. The Dudaim-types showed instead a rather more conservative behaviour, typical of isohydric plants, which privilege water loss avoidance even at low water restriction levels, at the cost of carbon assimilation and growth.

Selection of melon genotypes with resistance to <i>Didymella bryoniae</i> using a diallel approach

The development of melon lines resistant to gummy stem blight (GSB) is an important strategy for decreasing losses caused by this disease. Thus, selecting the best parents for such a goal is essential. We evaluated the general (GCA) and specific (SCA) combining abilities of lines and accessions to guide the selection of the most promising parents to develop GSB-resistant lines. Six genotypes representing two groups (group I - susceptible: group II - resistant) were crossed in a partial diallel mating design. The genotypes were evaluated for disease expression using a rating scale from 0 (resistant) to 4 (susceptible). Estimates of GCA were negative and significant for the accessions PI 420145 and PI 482398, which can be suggested for crosses aimed at obtaining melon lines resistant to GSB. There is a reciprocal effect for the resistance of melon genotypes to Didymella bryoniae, especially when PI 482398, PI 420145, and PI 140471 are crossed with JAB-20. Such results indicate that maternal effects may be involved in the resistance to gummy stem blight.

Stomatal and non-stomatal limitations are responsible in down-regulation of photosynthesis in melon plants grown under the saline condition: Application of carbon isotope discrimination as a reliable proxy

Salinity is one of the most severe environmental stresses limiting agricultural crop production worldwide. Photosynthesis is one of the main biochemical processes getting affected by such stress conditions. Here we investigated the stomatal and non-stomatal factors during photosynthesis in two Iranian melon genotypes “Ghobadlu” and “Suski-e-Sabz”, as well as the “Galia” F1 cultivar, with an insight into better understanding the physiological mechanisms involved in the response of melon plants to increasing salinity. After plants were established in the greenhouse, they were supplied with nutrient solutions containing three salinity levels (0, 50, or 100 mM NaCl) for 15 and 30 days. With increasing salinity, almost all of the measured traits (e.g. stomatal conductance, transpiration rate, internal to ambient CO2 concentration ratio (Ci/Ca), Rubisco and nitrate reductase activity, carbon isotope discrimination (Δ13C), chlorophyll content, relative water content (RWC), etc.) significantly decreased after 15 and 30 days of treatments. In contrast, the overall mean of water use efficiency (intrinsic and instantaneous WUE), leaf abscisic acid (ABA) and flavonol contents, as well as osmotic potential (ΨS), all increased remarkably with increasing stress, across all genotypes. In addition, notable correlations were found between Δ13C and leaf gas exchange parameters as well as most of the measured traits (e.g. leaf area, biomass, RWC, ΨS, etc.), encouraging the possibility of using Δ13C as an important proxy for indirect selection of melon genotypes with higher photosynthetic capacity and higher salinity tolerance. The overall results suggest that both stomatal and non-stomatal limitations play an important role in reduced photosynthesis rate in melon genotypes studied under NaCl stress. This conclusion is supported by the concurrently increased resistance to CO2 diffusion, and lower Rubisco activity under NaCl treatments at the two sampling dates, and this was revealed by the appearance of lower Ci/Ca ratios and lower Δ13C in the leaves of salt-treated plants.

Yield and Morphological İnvestıgatıon of Different Tigger Melon (Cucumis Melo Dudaim) Genotypes Belongıng to Sirnak/Turkey Region

This study was performed in 18 different tigger melon ( CUCUMİS MELO DUDAİM ) genotypes which were collected from the various villages of İdil/Sirnak. In the study, measurements of brix in fruit, SPAD, dry matter in fruit, total yield, seed weight, seed weight per fruit, seed length, number of seeds per fruit, fruit weight, fruit length, fruit width, fruit wall thickness, main stem diameter, main stem length, the main number of nodes in the stem, nod in the main stem were carried out. In addition, the experiment was performed with 4 replications including 10 plants per every replicate. The experiment was made according to the random design of the trial blocks. The data obtained at the end of the experiment statistical analyzes were made by evaluating with JMP8 program and the means were compared according to LSD test. In addition, cluster analysis was also performed in terms of similarity between genotypes in the experiment. As a result, when the fruit measurements are examined; total fruit yield was recorded with high amount in the tigger melon genotype, on the other hand, genotype number 13 of the tigger melon was recorded as the low. According to the obtained results, morphological and pomological measurements demonstrated difference from each other in 18 tigger melon melon plants and fruits. Keywords: Tigger melon, Seed, CUCUMİS MELO DUDAİM, SPAD, Brix. DOI : 10.7176/JBAH/9-10-09 Publication date :May 31 st 2019

Emergence and vegetative development of melon in function of the soil salinity

Soil salinity is one of the main factors limiting the development and global agricultural productivity. Melon (Cucumis melo L.) is mainly produced in arid and semi-arid regions around the world, favoring the accumulation of soluble salts in the soil. Therefore, this study aimed to evaluate the emergence and vegetative development of melon genotype under different levels of soil salinity. The assessments were performed based on the exchangeable sodium percentage (ESP) at 0, 20, 25, 30 and 35%. The emergence was affected around 30 percentage points, by comparing the control and the highest dose (35%). The emergence speed and the emergence speed index parameters were also affected in a similar way, as well as the length and dry matter of the shoot. The growth of the plants was also affected by the saline stress, being superior in the control (0%) in relation to the treatments with NaCl. The leaf area increased between weeks 4 and 7 after the planting and was slightly superior in the control in relation to the plants submitted to saline stress (20, 25, 30 and 35% of exchangeable sodium). The melon is more sensible to the saline stress in the seedling emergence than in the vegetative development over time. The development of melon seedlings is mainly affected in exchangeable sodium values superior to 20%. The analyzed hybrid is sensible to the saline stress even in 20% of exchangeable sodium.

REACTION OF MELON GENOTYPES TO THE ROOT´S ROT CAUSED BY Monosporascus

ABSTRACT Monosporascus root rot and vine decline (MRRVD) is an important disease in the melon crop in Brazil. In our research, we evaluated the reaction of 45 melon genotypes to M. cannonballus in two successive cropping seasons in a greenhouse. Melon seedlings were transplanted into naturally infested soil. The evaluation of the genotypes was performed after 55 days, using a score scale to group each genotype into one of five classes and to calculate the disease severity index. None of the genotypes showed an immune reaction to M. cannonballus, regardless of the season. In the first cycle, only 28.9% ('AF 464', 'Auraprince', 'DRG 2278', 'Estoril', 'Guaporé', 'Kousto', 'Mabel', 'Magisto', 'Massaï', 'Nectar', '8530', 'RZ 34130' and 'RZ 34292') had a similar reaction to the high resistance to the MRRVD. Most genotypes (66.7%) behaved as moderately resistant while 4.4% were susceptible and none were highly susceptible. Conversely, in the second cycle, no genotype showed a reaction similar to high resistance while 42.2% behaved as moderately resistant, 48.9% as susceptible, and 8.9% as highly susceptible. The severity levels ranged from 7.5 to 60% and 27.5 to 92.5% for the first and second cycles, respectively. The genotypes 'DRG 2278', 'Estoril', 'Guaporé', 'Kousto', 'Mabel', 'Massaï', 'RZ 34130' and 'RZ 34292' were identified as highly resistant and moderately resistant in the first and second cycles of cultivation, respectively. Therefore, these genotypes are promising sources of resistance to M. cannonballus and should be preferred in commercial fields infested with the pathogen.

Heterosis and Combining Ability of Melon Genotypes of Momordica Group

Heterosis and Combining Ability of Melon Genotypes of Momordica Group

Cucumber mosaic virus resistance and reproductive biology of brazilian melon accessions

Abstract In melon (Cucumis melo) the resistance to Cucumber Mosaic Virus (CMV) and the knowledge about reproductive characteristics are important for selection of genitors for breeding population. So, the objective was to investigate if different accessions of melon from a Brazilian germplasm bank have variability in resistance to CMV (FNY strain) and polymorphism in resistance loci markers, as well as whether there is genetic diversity on resistance and floral morphology descriptors and reproductive biology, aiming at genetic improvement. The symptoms induced by CMV were evaluated in 52 melon genotypes which were genotyped with three markers of single nucleotide polymorphisms related to CMV resistance. A sample of 24 genotypes was characterized for floral and reproductive descriptors. The significant variation in resistance levels allows classifying the accessions in five groups. The polymorphisms detected by the markers were independently distributed in these groups, consistent with the oligogenic quantitative expression of the melon’s resistance. There were significant differences between accessions in floral and reproductive descriptors. Through the Ward-MLM multivariate strategy, these accesses were distributed into seven groups with distinct reproductive characteristics, including variability in pollen viability. Therefore, there are Brazilian accessions of melon that have resistance to CMV, as well as variability in reproductive characteristics.

Elucidation of correlation and path coefficient analysis for various morphological attributes in elite genotypes of bitter melon (Momordica charantia L.)

Bitter gourd or bitter melon is undoubtedly one of the most significant members of the cucurbitaceous vegetable crops. Through the exploitation of this highly potential crop, novel mechanisms and strategies can be developed that will open the wide arrays for further improvement. Keeping this objective in mind, a field investigation was conducted with twenty genotypes of bitter gourd at Vegetable Research Farm of Banaras Hindu University to estimate the character association as well as direct and indirect effects of different yield traits on average yield per plant through correlation and path coefficient analysis. The data demonstrated that the positive and significant interrelationship with average yield per plant was obtained by various traits, viz., node number of first staminate flower (0.375), node number of first pistillate flower (0.54), vine length (0.417), fruit length (0.356), fruit circumference (0.79), average fruit weight (0.768), fruits per plant (0.637), and internodal length (0.3) while negative significant correlation was followed by days toanthesis of first staminate flower (-0.331) and days to first harvest (-0.355). The average fruit weight (0.572) had highest direct positive effect on average yield per plant followed by traits like days to fifty per cent flowering (0.296), number of fruits per plant (0.274), and number of primary branches per plant (0.259) whereas maximum direct negative effect was recorded by days to anthesis of first staminate flower (-0.263) followed by internodal length (-0.162) and days to first harvest (-0.152). The traits with high positive direct effect should be more prioritised for effective selection of promising genotypes for further crop improvement programmes.

Genetic diversity among melon ( Cucumis melo L.) accessions revealed by morphological traits and ISSR markers

The genetic relationships among 48 melon ( Cucumis melo L.) genotypes collected from various parts of Azerbaijan were determined by comparing their phenotypic and molecular traits. Eleven agromorphological traits and 10 polymorphic inter-simple sequence repeat (ISSR) primers were used to define the genetic diversity. Principal component analysis grouped the agromorphological traits into the first four axes, describing 78% of the total variations. The highest genetic variation coefficient was found for yield per hectare (20.32%) and for fruit length (17.35%). Calculated heritability for yield was 0.96. The analysis of morphological traits grouped the accessions into four clusters. The 10 ISSR primers yielded 35 polymorphic alleles, representing 85.4% of all the amplified loci. The average genetic diversity index determined was 0.70. The highest and the lowest similarity indexes were equal to 0.97 and 0.36, respectively. The 48 accessions were grouped into 10 clusters based on ISSR markers. Correlation between distance matrices based on agromorphological traits and ISSR markers was not statistically significant (r = 0.012).

Evaluation of powdery mildew resistance in various melon (Cucumis melo L.) genotypes

Evaluation of powdery mildew resistance in various melon (Cucumis melo L.) genotypes

Screening of melon genotypes identifies gummy stem blight resistance associated with Gsb1 resistant loci

Md Zahid Hassan, Arif Hasan Khan Robin, Md Abdur Rahim, Sathishkumar Natarajan, Hoy-Taek Kim, Jong-In Park, and Ill-Sup Nou. J Plant Biotechnol 2018;45:217-27. https://doi.org/10.5010/JPB.2018.45.3.217