Isozyme Banding Patterns Research Articles

Population study of Cucurbita moschata based on morphological characters and isozyme banding patterns in Bima District, West Nusa Tenggara, Indonesia

Abstract. Furqan M, Suranto, Sugiyarto. 2023. Population study of Cucurbita moschata based on morphological characters and isozyme banding patterns in Bima District, West Nusa Tenggara, Indonesia. Biodiversitas 25: 30-38. Pumpkin is a tropical plant that benefits from its lipid, protein, carbohydrate, and mineral contents. The morphological characters of pumpkins can be influenced by their genetics and environment. This research aimed to examine the relationship among 9 accessions of Cucurbita moschata Duchesne from 9 locations in Bima district, West Nusa Tenggara (NTB) province, Indonesia. The morphological characters of stems, leaves, and flowers were analyzed qualitatively and quantitatively and an ANOVA was used to look at the difference of leaf and flower characters. Polyacrylamide gel electrophoresis (PAGE) with peroxidase and esterase was used to detect isozyme banding patterns. Morphological characters and isozyme data were analyzed using a UPGMA method with NTSYS version 2.0 software to construct a dendrogram. The results show evidence of the varied morphological appearance of C. moschata from the different locations. A total of 12 esterase bands and 10 peroxidase bands were detected from Wera2. An undetected band with Rf value of 0.567 was recorded, while a unique band with Rf value of 0.133 (peroxidase) was not detected. Undetected bands with Rf values of 0.645 and 0.726 of esterase were also recorded. A dendrogram showed three clusters of 9 accessions studied, in which Wera2 and Wera3 each stood alone, and a third large group was formed from the 7 remaining accessions. These results confirm the uniqueness of the Wera2 and Wera3 accessions. These results would be good information in determining a management strategy for improving the quality of pumpkins in the future.

Electrophoretic banding patterns of esterase isozymes in fresh water fish Channa punctatus

Electrophoretic banding patterns of esterase isozymes in fresh water fish Channa punctatus

Isoenzyme Investigation and Morphometrics Study of Neckera complanata and Neckera crispa

Bryophytes are small, non-vascular, and non-flowering plants. Neckera complanata (Hedw.) Huebener and Neckera crispa Hedew.are one of the most prominent species in the Hyrcanian wetland found in glossy pale or yellowish-green patches. There is no evidence for morphometry and isoenzymes biochemical markers (Peroxidase/Superoxide dismutase) works on this genus in Iran. The purpose of this study is to investigate the differences and similarities among Neckera complanata and Neckera crispa moss populations in the north of Iran using morphometry and isoenzymes biochemical markers (Peroxidase / Superoxide dismutase). For this purpose, 18 populations from three provinces including Golestan, Mazandaran, and Guilan were collected at the same altitudes in autumn 2017. The results of morphometry were shown leaf length and leaf apex width/length were the most effective traits for the separation of populations. The results of the morphometry method and the results of isozyme banding patterns were the same, although very minor differences were observed. The largest diversity of Shannon (H) belongs to the population of Neckera crispa from Hezarjarib while other populations have low genetic diversity. Because of the destruction of many habitats in the northern provinces of Iran and the increase in pollution in these areas, it can be said as a general result that perhaps the reason for low genetic diversity in Neckera complanata and Neckera crispa populations is the gradual extinction of these two species.

Physiological and biochemical studies of black gram (Vigna mungo (L.) Hepper) under polyethylene glycol induced drought stress

Black gram (Vigna mungo L. Hepper) is an important nutritious legume plant, the loess plateau with strong drought tolerance and broad adaptability. To understand the adaptation mechanism of black gram under simulatedlevels of drought (5, 10 and 20% PEG) stress, and the growth characteristics, physiological parameters including levels of chlorophyll content, reactive oxygen species, and antioxidant enzymes were measured and the changes were compared with control. The result showed the root and shoot length, chlorophyll content, relative water content, stomatal activity, electrolyte leakage, antioxidant, and histochemical activity were carried out for treated and control plants. Experimentally, the drought-induced black gram seedlings of fresh weight, dry weight, and relative water contents found to be decreased with increasing concentrations of PEG. However, the electrolyte leakage and biochemical components (MDA content), antioxidant enzymes (SOD, CAT, POX), isozyme band patterns and histochemical detection of ROS accumulation (H2O2 and O2•–) level also enhanced under drought stress. The electrolyte leakage and MDA content were raised at 60% and 20% of severe PEG treatment. ROS accumulation was increased in the PEG treatment when compared to control. Overall, the rapid, accurate, and strong physiological observations from the electrolyte leakage and histochemical detection suggest that modification or enhancement of the biochemical and antioxidant contents would be determine the growth of black gram under PEG induced drought stress.

Isozyme banding patterns of drought tolerant sugarcane putative mutants

Genetic characterization based on isozyme is useful for plant breeding studies. Isozyme marker may be used to understand genetic variability of irradiated plant. The objective of this research was to recognise banding pattern variabilities of 34 putative irradiated mutants that survival drought screening, based on isozyme analysis. The research was conducted in Maret – April 2015 in Biology Laboratory, PAU IPB Bogor. Vertical polyacrylamide gel electrophoresis was used to analyse three enzymes: peroxidase (PER), Esterase (EST), Aspartate Aminotransferase (AAT). Isozyme analysis was carried out on gamma rays irradiated mutants of sugarcane “Kidang Kencana” that have been selected for drought tolerant. The result showed that there was a genetic variability on 34 putative mutant of drought tolerant sugarcane, that was shown by polymorphic banding patterns on PER and EST enzymes. AAT was monomorphic, no variation was shown amongs the mutants. PER enzyme had 17 banding patterns and EST enzyme had 7 banding patterns. Dendogram based on PER and EST on 34 putative mutants could be grouped into two clusters at 81.14% genetic similirity. The isozime analysis can be used as a specific identifier of a new mutant that will be developed as new varieties.

Characterization of some Cotton Genotypes Highly Resistant to Fusarium Wilt Disease by Electrophoresis of Peroxidase Isozymes and Seed Proteins.

Cluster analysis was used to evaluate the genetic diversity among 12 cotton genotypes highly resistant to Fusarium wilt disease. Cluster analysis was carried out based on electrophoretic banding patterns of peroxidase isozymes and seed proteins. Cluster analysis of peroxidase isozymes divided the genotypes into three distinct remotely related groups. Peroxidase isozymes were reliable to differentiate among genotypes included in the different groups; however, the isozymes were of limited reliability to differentiate among genotypes within groups. The genotypes were separated into a larg number of remotely related subclusters based on cluster analysis of their protein banding patterns. Therefore, it was easy to differentiate among the genotypes of the different subclusters. The present study demonstrated that cotton genotypes could be identified by their electrophoretic banding patterns of peroxidase isozymes and seed proteins. These results could be of practical value for cultivar identification and for seed purity tests.

Interpretasi Genetik Pola Pita Isozim pada Cherax quadricarinatus Hasil Budidaya di Purwokerto dan Bogor

A study on genetic interpretation of isozyme band patterns of C. quadricarinatus cultured in Purwokerto and Bogor showed the good visualization of aspartat transaminase, alkohol dehidrogenase, malat dehidrogenase, esterase and acid phosphatase. Twelve loci resulted low level of polymorphism (P= 0,25) and also low valve of heterozygosity (H= 0,09). It indicated that the genetic diversity of both populations was low.

Evidence of pollen features and peroxidase isozymes in their morphological complexity of ten local cultivars of sweet potato from Indonesia

Abstract. Suranto, Wahyuni D, Edy Purwanto E. 2019. Evidence of pollen features and peroxidase isozymes in their morphological complexity of ten local cultivars of sweet potato from Indonesia. Biodiversitas 20: 2511-2518. Sweet potato (Ipomoea batatas Lam) has been recorded to have many cultivars and variations in the morphological characters not only for the leaves but also for the tubers. Sweet potatoes had economically significant role, especially as an additional food due to its high content of carbohydrate and other nutrient values. In order to test whether the morphological variation in this species was also expressed at the micro character; such as pollen morphology and enzyme activity, ten sweet potato cultivars were used in this study. The morphological characters such as shape and structure of the pollen features were examined using Scanning Electron Microscopy (SEM). Accordingly, Polyacrylamide Gel Electrophoresis (PAGE) was used to look-at the banding pattern of peroxidase isozymes. Morphological characters of leaves, flowers, and tuber were observed qualitatively both for the typical shape and colors. More detail of flower shapes was drown manually especially their petal edges. The results showed that there was variation among the ten sweet potato cultivars confirmed not only on their morphological characters but also the isozyme banding patterns of ten cultivars tested. At least, two cultivar Antin 3 and Papua Solossa had uniques characters especially the purple stem and yellow tuber and dark skin on the tuber, respectively. In addition, the structural analysis of pollen showed that Beta 1 cultivar has very large pollen; 116,50 μm (axis polar) and the contour surface of exine pollen was concave with the aperture shape and ornamentation were prolate spheroidal and echinate respectively. This unique character was also shown by its peroxidase isozyme bands. Eleven bands of leaf and 13 bands of stem samples were detected respectively. The typical Rf values of 0.31, 0.48 and 0.41 were detected for the leaf, while for stem samples were 0.14, 0.20 and 0.26. The Beta 1 cultivar showed very thick bands in between Rf values of 0.13 and 0.26. Meanwhile, the unique band pattern of Papua Salosa was the absence of band at 0.38.

铬解毒机制诱导香根草的生长和抗氧化反应

This study investigated the chromium (Cr) detoxification mechanism-induced changes in growth and antioxidant defence enzyme activities in Chrysopogon zizanioides. Plant growth decreased by 36.8% and 45.0% in the shoots and roots, respectively, in the 50 mg/L Cr treatment. Cr accumulation was higher in root (9807 μg/g DW) than in shoots (8730 μg/g DW). Photosynthetic pigments and malondialdehyde content increased up to the 30 mg/L Cr treatment, whereas they declined at higher doses. The activity of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) were increased significantly with increasing of Cr dose but slightly declined at higher doses. Isozyme banding patterns revealed the expression of multiple bands for SOD, CAT and POX enzymes, and the band intensity decreased at high doses of Cr exposure. These results indicate that higher Cr doses increased the oxidative stress by over production of reactive oxygen species (ROS) in vetiver that had potential tolerance mechanism to Cr as evidenced by enhanced level of antioxidative enzymes, photosynthetic pigments, MDA contents. Therefore, vetiver has evolved a mechanism for detoxification and accumulates higher concentration of toxic Cr. This study provides a better understanding of how vetiver detoxifies Cr.

STUDI KOMPARASI KARAKTER MORFOLOGI DAN ISOZIM DUKU Lansium parasiticum (Osbeck) K.C. Sahni & Bennet ASAL KABUPATEN KUANTAN SINGINGI, RIAU

Duku ( Lansium parasiticum (Osbeck) K.C. Sahni & Bennet) from Kuantan Singingi Regency, Riau, known by local people as duku turak, has a unique oval betel nut-like neck, and tastes sweet similar to round duku. It has thick skin that is not easy to break, nor quickly rot. This duku was more preferred by local people than the round shaped duku known as duku gondok. However, the information about the potential of duku turak has not been known widely. The aim of this study was to compare morphology and isozyme characteristics of duku turak and gondok growing in Kuantan Singingi Regency based on morphological and isozyme characteristics. The result showed that turak and gondok can be distinguished based on morphological characteristics i.e tree, leave and fruit shape. Analysis of isozyme banding patterns used esterase isozyme which shows variation in the resulting patterns formed two bands. Acid phospatase (ACP) isozyme formed eight bands and aspartate aminotransferase (AAT) isozyme formed only one band. However, peroksidase isozyme showed no different variation in the resulting patterns. Similarity coefficient value based on morphology characters was about 0.15 – 0.74, while value of isozyme analysis was about 0.29 – 1.00 as well as the combination among morphology characters and isozyme were about 0.24 – 0.71. The results of this study provide characteristic information and taxonomic groupings that can be useful as a type improvement program that relies heavily on available genetic diversity

Morphological characteristics and isozyme banding patterns of Cucurbita moschata at different altitudes

Hidayati NR, Suranto, Sajidan. 2018. Morphological characteristics and isozyme banding patterns of Cucurbita moschata at different altitudes. Biodiversitas 19: 1683-1689. Aims of this research were to investigate the morphological character and isozyme banding patterns of Cucurbita moschata plants grown at three different altitudes. Samples in this study consisted of leaf, stem, and flowers. The morphological characters were conducted by direct observation in the field and analyzed descriptively as well as statically by one way ANOVA. The isozyme bands appearance of esterase and peroxidase of leaf samples were conducted using polyacrylamide gel electrophoresis (PAGE). Qualitative approach was used to analyze the presence and the absence of isozyme bands, while Retardation factor (Rf) was used to analyze quantitatively. The results showed that most plants grown at middle altitude (351-750 m asl.) were well-developed in terms of length of leaves, stems and flowers. Accordingly, the isozyme banding pattern of peroxidase was also found varied in plants grown at middle altitudes from which the presence of very unique bands was detected. Conversely, the band detected in plants grown at the lower and the highest altitudes was similar in term of band's number but it was different in the quality of the bands. Meanwhile, esterase isozyme banding pattern of plants grown at the lower and higher altitude had more bands than the middle altitude. Based on this result it is obvious that the isozyme data could be used to support in understanding the diversity morphological characters of plants grown in three different altitudes. This early result suggests that altitudes as a crucial factor in contributing the expression of isozyme appearance, which is useful for further pumpkin characterizations.

The Use of Electrophoretic Isozymes to Detect Tungro Infected Rice

Tungro is one of the most common diseases of rice plants which caused by double infection of RTBV (Rice Tungro Bacilliform Virus) and RTSV (Rice Tungro Spherical Virus), and it gives a significant economic loss. These viruses are transmitted by green leaf hopper ( Nephotteti x virescen s. Distant), and the interaction between host plant and the viruses were still quite difficult to be fully understood. In order to look at whether there are any differences between the healthy and tungro infected rice, this study was set to examine the character differences between the infected and the healthy plants based on isozyme banding pattern. The infected plants were collected from three districts around Surakarta (Sragen, Sukoharjo, Klaten) followed by Polyacrilamide Gel Electrophoresis (PAGE) to evaluate the isozyme banding patterns. There were peroxidase, esterase and acid phosphotase isozymes used. The results showed that the real differences of isozyme banding patterns of both healthy and tungro infected plants were discovered. In all cases, the Tungro infected rice had thicker and more band numbers compared to the healthy one. This evident suggested that Tungro infected rice could be detected early using PAGE method.

Effect of lead on phytotoxicity, growth, biochemical alterations and its role on genomic template stability in Sesbania grandiflora: A potential plant for phytoremediation

The present study was aimed at evaluating phytotoxicity of various concentrations of lead nitrate (0, 100, 200, 400, 600, 800 and 1000mgL−1) in Sesbania grandiflora. The seedling growth was significantly affected (46%) at 1000mgL−1 lead (Pb) treatment. Accumulation of Pb content was high in root (118mgg−1 dry weight) than in shoot (23mgg−1 dry weight). The level of photosynthetic pigment contents was gradually increased with increasing concentrations of Pb. Malondialdehyde (MDA) content increased in both the leaves as well as roots at 600mgL−1 Pb treatment and decreased at higher concentrations. The activity of antioxidative enzymes such as superoxide dismutase and peroxidase were positively correlated with Pb treatment while catalase and ascorbate peroxidase activities increased up to 600mgL−1 Pb treatment and then slightly decreased at higher concentrations. Isozyme banding pattern revealed the appearance of additional isoforms of superoxide dismutase and peroxidase in Pb treated leaf tissues. Isozyme band intensity was more consistent with the respective changes in antioxidative enzyme activities. Random amplified polymorphic DNA results indicated that genomic template stability (GTS) was significantly affected based on Pb concentrations. The present results suggest that higher concentrations of Pb enhanced the oxidative damage by over production of ROS in S. grandiflora that had potential tolerance mechanism to Pb as evidenced by increased level of photosynthetic pigments, MDA content, and the level of antioxidative enzymes. Retention of high levels of Pb in root indicated that S. grandiflora has potential for phytoextracting heavy metals by rhizofiltration.

Indonesia

Permatasari, Pitoyo A, Suratman. 2014. Keragaman suweg (Amorphophallus campanulatus) di wilayah eks karesidenan Surakarta berdasarkan karakter morfologi, anatomi dan pola pita isozim. Bioteknologi 11: 11-18. The aims of this research were to find out the diversity and relationship among elephant yam in Surakarta region based on variation of morphological, anatomical and isozyme banding patterns characters. Total 20 samples were taken using random purposive sampling technique in different altitude of 7 districts. Morphological, anatomical and isozyme banding pattern data were analyzed descriptively. The steady data of quantitative characteristic from morphological, anatomical and isozyme banding pattern of binary data was analyzed using an Index of Similarity (IS) and computed by the program Numerical Taxonomy and Multivariate Analysis System (NTSYS) version 2.10. The result showed that the elephant yam in Surakarta region were diverse based on morphological, anatomical and isozyme banding pattern, especially in ratio and color of petiolus; tuber ratio; index of stomatal; thick epidermis; thick mesophyll; term and ratio of palisade; term rafida; 4 peroxidase and 5 esterase isozyme banding pattern. Analysis of relationship of elephant yam based on morphological, anatomical and isozyme band pattern characters showed the similarity coefficient of 64,85% contained two major group. The first group consist of Boyolali 1, Boyolali 3, Boyolali 4, Wonogiri 4, Wonogiri 2, Karanganyar 1, Karanganyar 2, Karanganyar 3, Sragen 2, while the other samples were separated from this group.

ESTERASE ISOZYME PATTERNS DURING DEVELOPMENT OF BACTROCERA PAPAYA

One of the major horticultural crop pests is the Asian papaya fruit fly, Bactrocera papaya Drew & Hancock (Diptera: Tephritidae), in the tropical and subtropical regions of the world, which causes severe damage to the fleshy fruits and vegetables. Polyacrylamide gels were used for the first time to study the esterase isozyme patterns during different life stages and also to observe the enzyme patterns in the progeny (F1 generation) of B. papaya, which were maintained generation by generation under the laboratory condition. Four esterase isozymes, EST-1, EST-2, EST-3 and EST-4, were detected and their relative mobility values were 0.40, 0.26, 0.20 and 0.15, respectively. There was EST-1(0.40) with high mobility and EST-4(0.15) showed lowest mobility, closed to the cathode. These esterase isozymes were designated from the anodal end of the gel, i.e., the highest to lowest relative mobility. EST-1(0.40), EST-20.26 and EST-3(0.20) bands were only present in larval stage. EST-4(0.15) band was observed in all developmental stages except eggs. The thickness and the staining degree of bands varied in different developmental stages. In larvae, EST-1(0.40) band was very thin, EST-2(0.26) band was very thick and highly stained and EST-3(0.20) band was thin. The band of EST-4(0.15) was thick in larval and pupal stages, and comparatively very thick and highly stained in adults of both males and females. These variations of esterase isozymes indicated that it had stage-specific patterns in this pest species. In the progeny (F1 generation) of B. papaya, especially larvae and adults showed the similar esterase isozyme band patterns, comparing with their parents. The findings of the present research could be used for distinguishing this pest from other Bactrocera species as well as helpful to develop an environment friendly control method for this pest.

Effect of different light regimes on esterase isozyme profiles of three species of Drosophila

Circadian rhythm has been identified in every organism studied, from unicellular marine algae to man, and in virtually all physiological and biochemical functions. The endogenous circadian system functions to organize behaviour and physiology to adapt to and anticipate environment changes in light and temperature. The present study is an attempt to understand enzyme profiles (alpha and beta esterases) of Drosophila melanogaster (Oregon-K strain), Drosophila gangotrii and Drosophila jambulina under light/dark (LD), continuous light (LL) and continuous dark (DD) conditions over 30 generations. A polyacrylamide gel electrophoresis (7.5% – native gel) was used to study the esterase isozyme banding patterns in three species of Drosophila. It has been noticed that there were three alpha esterase loci in Drosophila species which were designated as α-est 1, α-est 2 and α-est 3. Similarly there were three beta esterase loci which were designated as β-est 1, β-est 2 and β-est 3. Flies maintained in different light regimes showed differences in their allelic patterns with respect to alpha and beta esterases. It was observed that there was expression of some bands at a given light regime and the absence of the same in another regime. This shows that the different light regimes affect the expression of esterase isozymes.

Isozyme Variation in Some Economically Important Species of the Genus Allium L. (Alliaceae)

Isozymes were used to study the phylogenetic relationships of selected species, varieties, and cultivars of Allium. Highest and lowest number of isozyme banding patterns was observed in Malate dehydrogenase and Malic enzyme, respectively. No intraspecific diversity was revealed within A. sativum and A. porrum. A. cepa showed some intraspecific diversity. Dendrogram showed close relationship among A. sativum and A. porrum. A. cepa was grouped with A. stracheyi and A. tuberosum. Principal coordinate analysis showed some differences with the dendrogram. This study showed that isozyme was more helpful for studying phylogenetic relationships at the species level rather than at intraspecific level.

Defense response of resistant and susceptible genotypes of castor (Ricinus communis L.) to wilt disease

The biochemical basis of resistance in castor (Ricinus communis L.) to Fusarium wilt, caused by the pathogen Fusarium oxysporum f. sp. ricini, was investigated. Induction of plant defence against pathogen attack is regulated by a complex network of different signals. Thus changes in various biochemical defenses including antioxidant enzymes, phenolic compounds and pathogenesis related (PR) proteins were investigated in the roots of resistant and susceptible genotypes of castor at 0, 24, 48 and 72 h.a.i. Infection by F. oxysporum significantly increased the superoxide dismutase (SOD) and peroxidase (POX) activities in the roots of susceptible genotypes, while the catalase (CAT) activities were appreciably higher in the roots of resistant genotypes at different stages. Constitutive levels of ascorbate peroxidase (APX) and polyphenol oxidase (PPO) were higher in the resistant genotypes. Also, the activities of phenylalanine ammonia lyase (PAL) and β 1, 3 glucanase significantly increased in the roots of the resistant genotypes after infections. The rate of increment of thiobarbituric acid reactive substances (TBARS) was higher in resistant genotypes after infection. Analysis of isozyme banding pattern of SOD, POX, PPO and esterase on native PAGE electrophoresis revealed that interaction between plant and fungi invoked various isozymes at 48 h of infection. SOD 3 was observed only in resistant genotypes at 24 h.a.i. except Geeta. Similarly induction of POX 5 was observed only in resistant genotypes at 48 h of infection, though the intensity of POX 5 was very less.

Paternity Analysis of Tea (Camellia sinensis L. Kuntz) Hybrids Using Isozyme Marker

Tea plant has been categorized as self-incompatible crop. This is the reason behind the high genetic diversity. Natural pollination is possible to occur and the male parent is usually unknown, therefore, there is a need of method to identify male parent of hybrids through paternity analysis. Isozyme markers have been successfully used for paternity analysis due to their co-dominant polymorphism. This research aimed to predict male parents of hybrids by figuring out the mating system through isozyme banding patterns. In this experiment, seven enzyme systems were evaluated, of which only two of the enzyme systems i.e. esterase and shikimate dehydrogenase showing clear band pattern of Est-1, Est-2, and Shd-1 loci. The mating system of tea could be categorized as a mixed mating model, with high estimated out-crossing rate of 98.6 %. The pollen contributors were not always originated from the vicinity of the female parents. Key words: isozyme markers, paternity analysis, tea

Characterisation of downy mildew resistant and susceptible pearl millet (Pennisetum glaucum (L.) R.Br.) genotypes using isozyme, protein, randomly amplified polymorphic DNA and inter-simple sequence repeat markers

In the present investigation, downy mildew resistant and susceptible pearl millet genotypes were characterised using seed protein and isozymes at pre- [45 days after sowing (DAS)] and post-infection (57 DAS, i.e. 7 days after infection) stage, as well as molecular markers at seedling stage without infection. Native polyacrylamide gel electrophoreis (PAGE) isozyme banding pattern of superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), ascorbate peroxidase (APX) and esterase showed some inducible band(s) due to disease infection and differentiated resistant and susceptible genotypes. Total seed protein profiling revealed the presence of two unique protein of ∼97 and ∼100 kDa in resistant genotypes. Randomly amplified polymorphic DNA (RAPD) analysis did not show any specific marker for disease resistant and susceptible genotypes. However, inter-simple sequence repeat (ISSR) markers showed six markers in resistant genotypes viz., UBC-825 (900 bp), UBC-827 (900 bp), UBC-857 (1000 bp, 700 bp, 375 bp and 200 bp). Moreover, a single unique band UBC-857 (400 bp) was present in only susceptible genotypes. Overall pooled analysis of isozymes, protein profiling, RAPD and ISSR data showed two distinct clusters of resistant and susceptible genotypes. These results suggested that seed protein profiling and ISSR markers may be used for large scale screening of germplasm for disease reaction trait.