Enzyme diversity in pearl millet (Pennisetum glaucum)

Summary The survey of enzyme polymorphism in West African pearl millet cultivars reported by Tostain et al. 1987 has been extended to include populations from other regions of Africa and from India. The eight enzyme systems studied included: alcohol dehydrogenase, -esterase, catalase, phosphoglucoisomerase, phosphoglucomutase, 6-phosphogluconate dehydrogenase, glutamate oxaloacetate transaminase, and malate dehydrogenase. One-hundred-ninety-nine populations of millet were analyzed, including 74 populations studied earlier. No new enzyme diversity was observed. Intrapopulation diversity ranged from 70%–90% of the total diversity, depending on their regions of origin. Four principal groups were distinguished in the following decreasing order of diversity: early-maturing cultivars from West and East Africa, late — maturing cultivars from West and East Africa, cultivars from India, and cultivars from southern Africa. The early-maturing cultivars were distributed between two principal focal points from East Africa in the East to Mali in the West. In the center were found millets from Niger which were most diverse. Indian and southern African cultivars were distinct, with the former appearing relatively similar to those of Niger, and the latter somewhat similar to late-maturing cultivars from West Africa, a diverse group that included late-maturing cultivars from East Africa. Based on the results obtained, an evolutionary hypothesis proposed here includes: multiple domestications in the Sahel, creation of early-maturing cultivars and their migration eastwards to India plus a southwards migration to Sudanian zone, and creation of late-maturing cultivars and their migration simultaneously westwards, eastwards, and southwards to southern Africa.     

肌酸激酶同工酶电泳对窒息新生儿心肌脑组织损伤的意义

目的:探讨血清肌酸激酶同工酶电泳对于新生儿窒息后发生心肌、脑组织损伤中的应用价值。方法:回顾性分析2009年1月至12月我院儿科ICU送检的CK同工酶电泳检测结果,并按不同疾病诊断进行分组统计。结果:在HIE、新生儿窒息患儿血清中CK-BB分别占总CK的13.9±16.8%、2.6±3.1%,CK-MB分别占总CK的2.5±2.0%、8.2±7.5%;CK-MM分别占总CK的91.4±14.1%、94.0±5.8%。新生儿窒息组可见CK-MB增加,显著高于对照及HIE组(P<0.01);HIE组患儿血清中可见CK-BB增加,与对照组、新生儿窒息组比较均有显著性差异(P<0.01)。结论:CK同工酶电泳对诊断新生儿窒息后引起的心脑损伤有重要价值。     

中国海菜花属植物三种同工酶的酶谱式样及其系统学含义

本文研究了代表中国海菜花属全部已知分类群的18个野生居群的过氧化物酶同工酶(POX),和其中7个居群的细胞色素氧化酶同工酶(COX),3个居群的酯酶同工酶酶谱式样及其系统学含义。结果表明:所有分类群的POX酶谱和龙舌草的COX酶谱均具多形现象,暗示编码有关同工酶的基因在种内居群间存在差异;3种同工酶都具分类价值,其分类结果与形态学分类结果一致支持将出水水菜花归并到水菜花中,取消海菜花的两个变种——路南海菜花和通海海菜花,而仅保留变种波叶海菜花。此外,本文初步揭示了POX酶谱式样与繁育系统的相关性。     

Peroxidases do not move in phloem

Phloem exudates from grafts between Queensland Blue pumpkin (Cucurbita maxima Duch.) and Candy Red Hawkesbury watermelon (Citrullus vulgaris Schrad.) were analysed by iso-electric focusing to detect iso-enzymes of peroxidase. These enzymes did not move in intact phloem but, when stems were cut, they surged rapidly through graft unions.     

An isoenzyme study in the genus Lotus (Fabaceae)

Summary Segregation of the cytosolic Pgi2 locus was studied among progeny of the synthetic allotetraploid (L. japonicus × L. alpinus)², the synthetic autotetraploid (L. alpinus)², and the cultivated tetraploid species L. corniculatus L. Evidence of an original diploid duplication found within the interspecific hybrid L. japonicus × L. alpinus was also found within the synthetic allotetraploid (quadruplication of loci). Evidence suggesting quadruplication of loci was also found in the tetraploid L. corniculatus, but not in the synthetic autotetraploid (L. alpinus)². It is suggested that the original duplication resulted from unequal crossing-over between homoeologues and that it provides evidence that L. corniculatus is a segmental allotetraploid. Quadruplication of loci in L. corniculatus could explain previously reported distorted tetrasomic ratios for segregation of qualitative characters in this species.     

Absence of association between heterozygosity and biomass production in Salix exigua Nutt

The relations between heterozygosity derived from a total of 12 variable isoenzyme loci and total above-ground leafless biomass production were examined in four full-sib families of Salix exigua, a willow species important in breeding efforts for short-rotation intensive-culture plantations. Relations were investigated by comparing the performance of heterozygotes with that of corresponding homozygotes in locus by locus comparisons, by investigating multiple regression models with individual loci as independent variables, and by employing the adaptive distance model. All these analytical approaches resulted in the manifestation of the absence of any relations between isoenzyme loci and biomass production. Possible reasons that may account for these results are discussed. Received: 8 October 1999 / Accepted: 4 November 1999     

The kinetics and thermal stability of phosphogluco-isomerase isozymes of ryegrasses (Lolium spp.)

The Michaelis constants (K_m) and activation energies (E_a) for allozymes of cytosolic phosphogluco-isomerase (PGI-2; EC 5.3.1.9) from ryegrasses ( Lolium perenne L., Lolium multiflorum Lam. and interspecific hybrids) have been investigated. Differences were found between the allelic isozymes, but intra-allelic variations were at least as large. The thermal stability of the isozymes also varied, with the most commonly occurring form (the b -allozyme) having the highest stability at 50°C. Some possible explanations for these findings are discussed and the implications for plant breeders outlined.     

Production and characterization of somatic hybrids between Solanum melongena L. and S. sisymbriifolium Lam.

Summary Protoplasts of 6-azauracil (AU) resistant cell lines of Solanum melongena L. were fused with protoplasts of S. sisymbriifolium Lam. to create somatic hybrids between these sexually-incompatible species. Following fusion, colonies were selected which were capable of growth in medium containing 1mM AU. These colonies were placed on medium containing zeatin which had been shown to stimulate anthocyanin production during shoot organogenesis in tissue explants of S. sisymbriifolium but not in S. melongena. A total of 37 anthocyanin-producing colonies were identified from which 26 hybrid plants were regenerated. The morphological traits intermediate to those of the parents included: flower colour, leaf shape, and trichome density. Cytogenetic analysis revealed that all hybrids were aneuploids but their chromosome numbers were close to the expected number of 48. Isozyme analysis revealed that nuclear genes of both parents were expressed in the hybrids. In addition, isoelectric focussing of the large subunit of ribulose 1,5-bisphosphate carboxylase (Rubisco) provided evidence that each hybrid expressed only the S. sisymbriifolium chloroplast genome. All hybrids regenerated thus far have been sterile.Contribution No. 787 Ottawa Research Station     

Hymenolepis microstoma: lactate and malate dehydrogenases of the adult worm.

Lactate and malate dehydrogenases (EC 1.1.1.27 and EC 1.1.1.37, respectively) were precipitated with ammonium sulfate, redissolved in 100 mM phosphate buffer, and the kinetic parameters of each enzyme determined. Lactate dehydrogenase: The enzyme preparation had a specific activity of 0.35 μmole NADH oxidized/min/mg protein for pyruvate reduction, and 0.10 μmole NAD reduced/min/mg protein for lactate oxidation. K_m values for the substrates and cofactors were as follows: pyruvate = 0.51, mM; lactate = 3.8 mM; NADH = 0.011 mM; and NAD = 0.17 mM. NADPH, NADP, or d(?)-lactate would not replace NADH, NAD, or l(+)-lactate, respectively. The enzyme was relatively stable at 50 C for 45 min, but much less stable at 60 C; repeated freezing and thawing of the enzyme preparation had little effect on LDH activity. Both p-chloromercuribenzoate (p-CMB) and N-ethylmaleimide (NEM) significantly inhibited LDH activity. Polyacrylamide gel electrophoresis demonstrated the presence of at least two LDH isoenzymes in the unpurified enzyme preparation. The molecular weight was estimated at 160,000 by gel chromatography. Malate dehydrogenase: The enzyme preparation had a specific activity of 6.70 μmole NADH oxidized/min/mg protein for oxaloacetate reduction, and 0.52 μmole NAD reduced/ min/mg protein for malate oxidation. K_m values for substrates and cofactors were as follows: l-malate = 1.09 mM; oxaloacetate = 0.0059 mM; NADH = 0.017 mM; and NAD = 0.180 mM. NADP and NADPH would not replace NAD and NADH, respectively, d-malate was oxidized slowly when present in high concentrations (>100 mM). Significant substrate inhibition occurred with concentrations of l-malate and oxaloacetate above 40 mM and 0.5 mM, respectively. The enzyme was unstable at temperatures above 40 C, but repeated freezing and thawing of the enzyme preparation had little effect on MDH activity. Only p-CMB inhibited MDH activity. Polyacrylamide gel electrophoresis demonstrated the presence of at least three MDH isoenzymes in the unpurified enzyme preparation, and the molecular weight was estimated at 49,000 by gel chromatography.     

Characterization of Two Cytosolic Diacylglycerol Kinase Forms

Abstract: Two forms of rat brain cytosolic diacylglycerol kinase (EC 2.7.1.107) were separated by heparin-agarose column chromatography. These forms, designated DGK-I and DGK-II, were not interconvertible as determined by rechromatography. DGK-I and DGK-II had respective molecular masses of 88 and 180 kDa, as measured by Sepharose 6B chromatography. Both forms preferred diacylglycerol over monoacylglycerol and were insensitive to R59022. DGK-II, but not DGK-I, was activated by an activator substance prepared from chicken egg yolk. DGK-II was activated by a rat brain cytosolic activator and was exclusively sensitive to 5'-AMP-mediated inactivation. Further studies revealed that these two forms had the following distinct characteristics: (a) substrate specificity, (b) inhibition by heparin, (c) sensitivity to lysine-containing polyamino acids, and (d) responses to different phospholipids. In general, DGK-II was more responsive to various inhibitors and activators, making it a prime candidate for a regulatable enzyme.