Leaf Esterase Research Articles

Changes of cotton leaf worm haemocytes and esterases after exposure to compounds derived from urea and Rice Straw

In the current study, the role of cotton leaf worm haemocytes and esterases in detoxification were recorded. The haemolymph was collected from the 6 th instar larvae which treated with LC25, LC50, LC70 of three newly compounds extracted from wastes from natural origin, Cyano acetyl urea (CAU), Benzimidazolyl acetyl urea (BAU) from urea and Cyano acetyl urea (CAH) from rice straw as 4 th instar larvae. The mean of total haemocytes counts in haemolymph of untreated 6 th larval instar was 9430±35.78 cells/mm 3 . Data indicated an increase in the total haemocytes of treated ones with CAU, at LC25 and LC50, BAU at LC25 and CAH at LC25, LC50 of CAH the percentages of change were: 19.01, 14.31, 1.90, 21.00 and 14.55, respectively. In addition, the percentages of oenocytoid counts were increased in all concentrations for all compounds except for CAH (at LC70) as it did not change comparing control count. On contrast, data cleared reductions in granulocytes in all concentrations for all compounds except for BAU at LC50 as they increased slightly (15.12%). On the other hand, the results were showed great differences in number of zones of esterase activity and in substrate specificity between treated and untreated samples. For instance, in the case of α - naphthyl acetate, 36 esterase bands were detected in control and treated larval samples with Rf ranging between 0.01 to 0.28. While, in the case of  - naphthyl acetate, 39 esterase bands were detected in control and treated larval samples with Rf ranging between 0.01 to 0.92.

Allozyme variations in leaf esterase and root peroxidase isozymes and linkage with dwarfing genes in induced dwarf mutants of grass pea (Lathyrus sativus L.)

Genetic basis of different leaf esterase and root peroxidase isozymes was investigated by analyzing their zymogram phenotypes in selfed and intercrossed progenies of two local varieties (used as control) and three induced true breeding dwarf mutant lines of grass pea (Lathyrus sativus L.). Two non-allelic genes, df1/df2 and df3 controlling dwarfism in grass pea were included in the present linkage studies with different isozyme loci. The dwarf mutants could be distinguished from one another and also from control varieties by the presence of unique allozyme/s coded by allele/s in different loci. A good fit to 1: 2: 1 in F2 and 1:1 in back cross indicated monogenic segregation of isozyme loci with co-dominant expression of different alleles and the isozymes were functionally monomeric. Linkage analysis revealed closely linked associations among Est-1, Est-2and df3 loci and also between Est-3 and Prx-1 loci. All five genes, however, assorted independently with df1/df2 as well as with prx-3. Locus prx-2 was monomorphic in all five parents. Key words: Dwarf mutants, isozymes, linkage, Lathyrus sativus L.

Isozymes variability among Fusarium udum resistant cultivars of pigeonpea (Cajanus cajan (L.) (Millsp)

The present study was carried out to select the different pigeonpea cultivars for resistance against wilt caused by Fusarium udum and to assess the genetic variability among the resistant and susceptible cultivars. These cultivars were screened by root dip inoculation and classified into resistant (ICP 8863 and 9145), moderately resistant (ICP 11681 and Selection-1), susceptible (ICP 7118, TRG-1 and LRG-30) and highly susceptible cultivars (ICP-2376 and LRG-41). The peroxidase activity (PEO) in both leaf and root tissues of four pigeonpea cultivars (ICP 8863, Selection-1, ICP 2376 and LRG-30) were determined at 1st, 4th and 7th day after inoculation (DAI) in healthy and F. udum infected tissues. Higher PEO activity in both leaf and root was observed and at 4th DAI in susceptible cultivars. In native-PAGE analysis of isozymes, the induction of specific leaf peroxidase band (Em=0.17) and two root peroxidase bands (Em=0.24 and 0.55) were observed in ICP 8863 after inoculation. Significant differences were observed in the leaf phosphatase and esterase banding profiles of all the cultivars. The presence of leaf phosphatase band at Em of 0.04 was observed only in ICP 8863 and 11681. The leaf esterase band (Em=0.3) was well expressed in ICP 8863 when compared to other cultivars. The significance of peroxidase in plant defense mechanism and utility of biochemical markers in breeding programmes are discussed.

Chromosomal location of an esterase gene set (Est-10) of common wheat orthologous to Est5 of Aegilops tauschii

Est5 is a monomeric, polymorphic, electrophoretically slow leaf esterase of Aegilops tauschii. Previously it was found that Est5 was involved in the process of adaptive intraspecies divergence, and was genetically linked with another Ae. tauschii enzyme locus of unknown chromosomal location — Nadhd2. The genes orthologous to Est5 are found to be located in chromosomes 3DL and 3AL of common wheat cultivar ‘Chinese Spring’.

Caracterização isoenzimática de genótipos de arroz

Objetivou-se, neste trabalho, caracterizar isoenzimaticamente genótipos de arroz (Oryza sativa L.). A produtividade do arroz irrigado no Rio Grande do Sul é elevada, em virtude da alta tecnologia e sistema de irrigação usados, associados ao potencial alcançado pelas cultivares desenvolvidas através de melhoramento genético. Apenas seis ancestrais contribuem com 86% dos genes das cultivares mais plantadas. Como conseqüência desta estreita base genética, as cultivares apresentam um alto grau de parentesco e de similaridade de suas características morfológicas e agronômicas, o que dificulta a identificação varietal. A concorrência com genótipos, como arroz-vermelho e arroz-preto, de difícil controle por serem da mesma espécie que os cultivados, é considerada como um dos maiores problemas da cultura. Análises de isoenzimas podem ser usadas para o estudo da variabilidade e para estimar as relações genéticas existentes entre estes genótipos. Eletroforese em gel de poliacrilamida foi empregada, portanto, para caracterizar, através de isoenzimas de esterase, 6fosfogluconato desidrogenase, fosfoglucoisomerase e isocitrato desidrogenase em sementes e folhas de plântulas, e de fosfatase ácida e aspartato transaminase em folhas de plântulas, as cultivares BR-IRGA 409, BR-IRGA 410, BRS 6 ('Chuí'), BRS 7 ('Taim'), BRS Agrisul, INIA Taquari, El Paso L 144 e IRGA 417, e ecótipos de arroz-vermelho e arroz-preto. A análise de agrupamento, efetuada por meio do coeficiente de Jaccard e pelo método da média aritmética não ponderada (UPGMA), possibilitou a diferenciação de todos os genótipos, à exceção de BRS 6 ('Chuí') e BRS 7 ('Taim'). Três grupos foram identificados, incluindo-se, em um deles, os ecótipos de arroz-vermelho e arroz-preto, que apresentaram 95% de similaridade.

Erucastrum abyssinicum Brassica oleracea hybrids obtained by ovary and ovule culture

Pollen germination and pollen tube growth were studied in the cross Erucastrum abyssinicum × Brassica oleracea. 25 per cent ovules showed pollen tube entry into the micropyle 5 days after pollination. Embryos did not grow beyond the globular or heart-shaped stage. Thus post-fertilization barriers prevent hybridization in this cross. Sequential culture of ovaries and ovules 10 days after pollination helped rescue globular embryos. Three hybrids were obtained. All the hybrids flowered in vitro. Pollen fertility was 6.5 per cent. Leaf esterase and peroxidase isozymes confirmed the hybrid nature. Meiotic metaphase in the pollen mother cells showed 25 chromosomes. The presence of up to 9 bivalents and a quadrivalent indicates high homology between the two genera. The intergeneric hybrids have been multiplied using nodal segment culture. They are a source for obtaining cms cole crops and widening the gene pool of B. oleracea.

Variation of leaf esterases in some Ethiopian tetraploid wheat landraces

Esterase isozymes of tetraploid wheat landraces originating from four localities in the central highlands of Ethiopia were studied using the horizontal starch gel electrophoresis. The landraces studied consisted of 60 Triticum turgidum L., one T. dicoccon Schrank, one T. polonicum L. and an improved local cultivar. One introduced durum cultivar was also included for comparison.

Mapping of genes for copper efficiency in rye and the relationship between copper and iron efficiency

A 4B/5R wheat-rye translocation line derived from the Danish wheat variety ‘Viking’ was revealed to be highly copper efficient. The chromosomal exchange includes a very small terminal segment of chromosome arm 5RL of rye which was physically mapped by genomic DNA: DNA in situ hybridization and chromosome analysis. The gene for Cu efficiency (Ce) is linked to a dominant hairy neck character from rye (Ha1) and to two rye-specific leaf esterase loci (Est6, Est7), all of which are postulated to map to the distal part of 5RL. Genes coding for mugineic acid synthetase and 3-hydroxymugineic acid synthetase also on chromosome 5R are not included in the 4B/5R translocation and hence map outside the terminal 5R region. These genetic and molecular markers can be useful tools for large-scale screening in wheat breeding programmes.

A proposed revision of the IBPGR barley descriptor list

The International Board for Plant Genetic Resources (IBPGR) promote a set of morphological characters considered to be the minimum number for the description of crop germ plasm collections. The format and style of these IBPGR minimum descriptors are typical across numerous crop and species genera. However, no research publications have reviewed the relative usefulness of individual descriptors, nor indeed if additional descriptors should be included into the "minimum descriptor list".A diverse collection of 1,379 spring sown barleys were evaluated for the 12 IBPGR "minimum" descriptors, an "additional" 9 morphological descriptors of similar type of the minimum descriptors, and a contrasting group of 3 hordein and 3 leaf esterase biochemical markers. Multivariate analyses were performed on combinations of the "minimum", "additional" and biochemical markers to reveal detailed associations amongst these various descriptors. The IBPGR minimum descriptor list for barley was shown to be inadequate in descriminating between accessions, and a revised list is proposed.

Intergeneric somatic hybridization in Gramineae: somatic hybrid plants between tall fescue (Festuca arundinacea Schreb.) and Italian ryegrass (Lolium multiflorum Lam.)

Tall fescue (Festuca arundinacea Schreb.) protoplasts, inactivated by iodoacetamide, and non-morphogenic Italian ryegrass (Lolium multiflorum Lam.) protoplasts, both derived from suspension cultures, were electrofused and putative somatic hybrid plants were recovered. Two different genotypic fusion combinations were carried out and several green plants were regenerated in one of them. With respect to plant habitus, leaf and inflorescence morphology, the regenerants had phenotypes intermediate between those of the parents. Southern hybridization analysis using a rice ribosomal DNA probe revealed that the regenerants contained both tall fescue- and Italian ryegrass-specific-DNA fragments. A cloned Italian ryegrass-specific interspersed DNA probe hybridized to total genomic DNA from Italian ryegrass and from the green regenerated somatic hybrid plants but not to tall fescue. Chromosome counts and zymograms of leaf esterases suggested nuclear genome instability of the somatic hybrid plants analyzed. Four mitochondrial probes and one chloroplast DNA probe were used in Southern hybridization experiments to analyze the organellar composition of the somatic hybrids obtained. The somatic hybrid plants analyzed showed tall fescue, additive or novel mtDNA patterns when hybridized with different mitochondrial gene-specific probes, while corresponding analysis using a chloroplast gene-specific probe revealed in all cases the tall fescue hybridization profile. Independently regenerated F. arundinacea (+) L. multiflorum somatic hybrid plants were successfully transferred to soil and grown to maturity, representing the first flowering intergeneric somatic hybrids recovered in Gramineae.

Confirmation of a 4BL/5RL Wheat‐Rye Chromosome Translocation Line in the Wheat Cultivar ‘Viking’ Showing High Copper Efficiency

AbstractThe N‐ and C‐banding patterns of the chromosomes of the hexaploid wheat cultivar ‘Viking’, and the hairy‐necked line derived from it, were described. Except for chromosome 4B (formerly 4A), no significant variation in banding patterns were found between the two genotypes. In addition to a 5BS.7BS (5BL.7BL) translocation in both genotypes, the presence of a small 5RL rye segment in a terminal position on the long arm of chromosome 4B was suggested by banding and confirmed by meiotic analysis, and by the hairy neck marker as well as by rye‐specific leaf esterase bands in the translocation line. The translocation breakpoint is presumed to be located in the distal 4BL2.2 and/or 5RL2.3 band. The hairy‐necked ‘Viking’ was shown to be copper efficient. Its utilization in wheat breeding programmes is proposed.

Degree of apomixis in Chinese chive (Allium tuberosum) estimated by esterase isozyme analysis.

The degree of apomixis of six cultivars of Chinese chive (Allium tuberosum ROTTL.) (2n= 32, 4x) was estimated by electrophoretic analysis of leaf esterase (EST) in progenies obtained from crosses between these cultivars. In each progeny, more than 90% of the seedlings showed the same zymogram as their pistillate parent, the remaining seedlings showing hybrid zymograms. The differences between maternal and hybrid zymograms were clear and unmistakable. The degree of apomixis was therefore calculated to be higher than 90% for each cultivar tested. For two progeny arrays the results of the electrophoretic analysis were supported by simultaneous examination of two morphological traits, namely, outer tepal length and first flowering month. Among the seedlings showing the maternal EST zymogram, two dihaploids (2n= 17, 16), one each in two progenies, were detected by cytological examination of root tips. Those dihaploids being eliminated, the degree of apomixis estimated by the isozyrne analysis was reduced by 1% resp. 2% for these progenies. This modification, however, was of little significance because of the infrequent occurrence of dihaploids. The present results indicate that a progeny test exclusively based on the EST analysis gives only a slight overestimation of the degree of recurrent apomixis in Chinese chive and that Chinese chive is a facultative apomict wlth a high degree of apomixis.

Est-7, a set of genes controlling green tissue esterases in wheat and related species

Analysis using isoelectric focusing of "Chinese Spring" wheat genetic stocks revealed a set of coleoptile and leaf esterase loci, designated Est-7, on the long arms of the group 2 chromosomes. A survey of 38 other hexaploid genotypes revealed onyl a single variant, at Est-D7. Homoeoloci were found on chromosome (arm) 2HL of Hordeum vulgare, 2RL of Secale cereale, 2R (m)α of S. montanum, 2U of Aegilops umbellulata, 2E of Agropyron elongation and 2V of Dasypyrum villosa.

Genetic variability of leaf esterases in Triticum aestivum L. 2n=6x=42

Starch gel electrophoresis with two different buffer systems and several substrates and inhibitors have been used to study the electrophoretic variability of esterases in leaves of cultivars of Triticum aestivum. Each one of the buffer systems showed different levels of variability, according to the electrophoretic patterns. At the same time green and etiolated leaves showed different patterns in each buffer system. The variability was dependent upon the developmental stage of the leaves. According to the results from chromosomal location, the genes controlling esterases in green leaves were located in homoeology group 3, while the genes controlling esterases in etiolated leaves were in homoeology group 6. But both esterase isozymes showed a similar electrophoretic migration and a similar reponse to substrates and inhibitors. The possible origin of both sets of genes due to an interchromosomal duplication is discussed.

Genetic control of esterase-6 isozymes in hexaploid wheat and rye

The EST-6 leaf esterase phenotypes from euploid, nullisomic-tetrasomic and rye chromosome addition and substitution lines of common wheat were determined using polyacrylamide gel electrophoresis. Evidence is presented to demonstrate that Est-6 is a new set of genes, that are expressed in the leaf. The Est-6 gene set were clearly distinguished from the Est-5 genes which are expressed in the grain. The three homoeoallelic loci, Est-A6, Est-B6 and Est-D6, were located on chromosomes 3A, 3B and 3D. An Est-R6 gene was located on chromosome 6R is involved in rye. Some considerations concerning homoeology between homoeologous group 3 of wheat and the rye chromosome 6R are made.

Ottelia emersa Z.C. Zhao et R.L. Luo (hydrocharitaceae) — a Synonym of O. cordata (Wallich) Dandy

Based on observations of wild populations of Ottelia emersa Z.C. Zhao et R.L. Luo and O. cordata (Wallich) Dandy, cultivation experiments and electrophoresis of three enzymes in the leaves in these taxa, we have found that: (1) heterophylly is related to the age of the plants involved; ( (2) the developmental process and morphology of the leaves is identical in both taxa; (3) seed hair density and colour show continuous variation; (4) band patterns of the leaf esterase and cytochrome oxidase are identical, and that of peroxidase is substantially the same in the two taxa. Therefore we conclude that O. emersa and O. cordata are conspecific, and the former is a synonym of the latter.

Species relationship in the Pennisetum gene pool: enzyme polymorphism

Variation in leaf esterases (EST), 6-phosphogluconate dehydrogenase (PGD), shikimate dehydrogenase (SKDH), leucine aminopeptidase (AMP), phosphoglucomutase (PGM) and malate dehydrogenase (MDH) is reported in the Pennisetum gene pool. In the primary gene pool, polymorphism for EST, AMP, SKDH was very high, as compared to the near-monomorphic isozymes of PGD. Two loci controlling leaf esterases Est-1 and Est-2, were identified in the primary gene pool. Differences in allelic frequency distribution of the polymorphic Est-1 locus occur between the cultivated and wild pearl millet. The prevalent alleles of Est-1 are absent in P. purpureum Schumach (secondary gene pool). A monomorphic band of the α-esterase-specific Est-2 locus was identified in most of the secondary gene pool accessions, P. squamulatum Fresen and an accession of P. pedicellatum. SKDH and EST revealed differences between most of the tertiary gene pool species. By contrast, a PGD zymogram was prevalent in several species of different sectional taxa. Gene duplication for PGD isozymes occurs in the diploid species, P. ramosum, of the tertiary gene pool. Heterodimers of PGD and EST were observed in the hybrid between pearl millet and P. squamulatum, whereas a monomeric structure characterized SKDH and AMP.

Wide hybridization in Brassica

Crosses were made to obtain interspecific hybrids between B. fruticulosa (wild species ♀, 2n = 16) × B. campestris (cultivar ♂, 2n = 20). Although many pollen grains germinated and their tubes entered the style, only about 30% of the ovules received pollen tubes. Fertilized ovules aborted at various stages of development. A few hybrid seeds resulted from hand pollinations in the field, and they showed poor germination and seedling establishment. The in vitro culture of ovaries, ovules, and seeds increased the frequency of obtaining hybrid seeds and plants: the most effective method was ovary culture followed by ovule culture. The hybrid nature of the plants was confirmed through morphological, cytological, and electrophoretic studies. A meiotic analysis of F1 hybrids (2n = 18) showed that they had 0–5 bivalents and were completely pollen sterile. Electrophoretic analysis of leaf esterases and acid phosphatases of F1 hybrids revealed bands derived from each parent. Induced amphidiploids of F1 hybrids contained mostly bivalents, and had about 50% fertile pollen.

A selection method for the synthesis of triploid hybrids by fusion of microspore protoplasts (n) with somatic cell protoplasts (2n)

Microspore protoplasts (n) isolated at the tetrad stage from plants of Nicotiana tabacum Km(+) (2n=4x=48) were fused with somatic cell protoplasts (2n) of WT N. rustica (2n=4x=48) to produce triploid plants. A total of 21.2×10(6) microspore protoplasts were fused with 11.2×10(6) somatic cell protoplasts using the high pH/Ca(+ +) method. Microspore protoplasts did not divide and WT N. rustica protoplasts stopped dividing when the protoplast-derived colonies were transferred to a selection medium containing kanamycin. A total of 104 actively growing green colonies were recovered on the selection medium. Ninety-six of these colonies were tested for their hybrid nature by PAGE of peroxidases and were found to contain bands characteristic of both parents. Hybrid nature of the plants regenerated from some of the selected colonies was confirmed by IEF of leaf esterases, by NPT II activity assay and by hybridizing total DNAs restricted with EcoR I to a cloned 18s rDNA fragment. Root tip squashes of six of the hybrid plants revealed chromosome numbers ranging from 58-72. From chromosomal and biochemical analysis, it can be concluded that the procedure of fusing microspore protoplasts (n) of species A carrying a dominant selection marker with WT somatic cell protoplasts (2n) of species B can be a convenient selection method for the synthesis of triploid plants. The significance of triploids lies in their subsequent use for transferring alien chromosomes and genes of species A to species B.

Character inheritance and polymorphism in a wild oat (Avena fatua) population

A population of wild oats (Avena fatua L.) in a cultivated field was sampled once or twice a year between 1979 and 1983. Plants differing in seed morphology, seed avenin pattern, leaf esterase pattern, and vernalization requirements were crossed to study the inheritance of these traits (13 loci). The polymorphism of the population for each of these characters and 11 others was investigated as well as the frequency of each type of plant (phenotype). Forty-seven phenotypes were observed over the 5 years, but only five were consistently present in all samples. They represented 78% on average of the population. The observed increase of a phenotype correlatively to the decrease of another one is discussed as a function of cultural practices.