从番茄过氧化物酶同工酶表型差异探讨杂种优势机理

用聚丙烯酰胺凝胶电泳对番茄进行过氧化物酶同工酶分析表明:(1)不同栽培品种在同一生育期或同一品种在不同生育期、不同部位酶谱表型有明显差异,但把各品种不同生育期的酶谱叠加,所得到的总酶谱是相同的;(2)品种间杂交其F_1的过氧化物酶同工酶谱不存在“杂种酶带”或“互补酶带”;(3)品种间过氧化物酶同工酶的结构基因相同,酶谱表型上的差异可能是基因表达顺序上的差异,这可能是番茄杂种优势产生的生理基础之一。选择早期(芽期)酶谱表型差异适中的品种作杂交亲本,有可能获得生产上可用的高优组合。     

荞麦属植物三叶期幼叶过氧化物酶同工酶研究

用聚丙烯酰胺凝胶电泳技术对荞麦属(Fagopyrum Mill)8个种(含大粒组7个种和小粒组1个种)28份栽培及野生荞麦植株三叶期幼叶的过氧化物酶同工酶进行了研究.结果发现:过氧化物酶同工酶酶带共23条,不同物种的酶带数4～9条,其中甜荞有6条带,而苦荞为9条.酶带及聚类分析表明:大粒组荞麦种的谱带与细野荞(F.gracilipes)等小粒组荞麦种间差异极大,甜荞(F.esculentum)和苦荞(F.tataricum)酶带分别与大野荞(F.megaspartanium)和毛野荞(F.pilus)相似,并分别与大野荞和毛野荞聚类最近, 支持Chen(1999)提出的大野荞和毛野荞可能分别是甜荞和苦荞的祖先种的假说.     

马铃薯叶甲属六个种的等位基因酶变异

用聚丙烯酰胺凝胶电泳测定了马铃薯叶甲属(Leptinotarsa)六个种:马铃薯叶甲(L. decemlinecta)、胡颓子叶茄叶甲(L. texana)、柔毛茄叶甲(L. rubiginosa)、膜苞菊叶甲(L. lineolata)、蒺藜叶甲(L. peninsularis)和蒺藜四条叶甲(L. tlascalana)的等位基因酶变异.分析了14种同工酶(ACPH、ADH、AMY、FUM、GDH、GOT、IDH、HBDH、LAP、MDH、ME、PGM、SORDH和XDH)的17个座位.其中单型座位有9个,多型座位有8个,后者占47.1%.六个种的平均杂合度为0.091.其中最高的是膜苞菊叶甲0.154;最低的是蒺藜四条叶甲0.042.六个种彼此之间的遗传距(D)在0.129—0.573之间,说明这六个种的亲缘关系是很密切的.     

天麻3种变型过氧化物酶的同工酶研究

采用聚丙烯酰胺凝胶电泳（PAGE）对天麻3种变型的过氧化物酶（POD）的同工酶进行了研究。结果表明,天麻3种变型的过氧化物酶的同工酶活性不同,酶谱条带数在5～7条之间,且具有特征谱带。其中乌天麻和红天麻的酶谱条带数相同,两者的酶谱条带数均多于绿天麻;绿天麻、乌天麻和红天麻的酶谱条数依次分别为5条、7条和7条;其中基本酶带有5条,Rf值分别为0．06、0．24、0．83,0．89、0．98;乌天麻与红天麻的相似度指数为0．86,说明这两个种亲缘关系近;乌天麻与绿天麻的相似度指数为0．71,反映它们之间的亲缘关系相对较远。     

赖草属七个种同工酶研究

用聚丙酰胺凝胶电泳法对赖草属７个种的幼根、幼叶进行了酯酶、过氧化物酶同工酶分析,结果表明：无论从相同器官进行不同的同工酶分析,还是从不同器官进行相同的同工酶分析,这７个种的酶谱在各带区存在相似酶带,但更多的是相异酶带。从酯酶、过氧化物酶这两种酶分析结果来看,酯酶比过氧化物酶分离效果好些;从幼根和幼叶这两个器官的酶谱来看,幼根比幼叶酶带多些,分离效果也好些。同时也表明这７个种的酶谱变化与染色体倍性变化无关     

Cucumis属3种不同倍性种间杂交后代的同工酶分析

以3种不同倍性的Cucum is属种间杂交后代[异源四倍体C.hy tivus(2n=4x=38,HHCC)、异源三倍体(2n=3x=26,HCC)和正反交种间杂种F1(2n=2x=19,HC/CH)]及其双亲为试材,比较研究了过氧化物酶(POD)和酯酶(EST)同工酶在不同器官中的酶谱表达特性.结果表明,花蕾中的POD和EST同工酶谱带都比叶片中的丰富,表现出明显的组织特异性.在相同器官的同一酶系统中,3种不同倍性种间杂交后代的酶谱基本一致,主要表现为互补双亲的酶带,同时出现了双亲所没有的酶带(POD4c和EST3b等),表明远缘杂交扩大了黄瓜的遗传基础.此外,在幼叶和花蕾的POD同工酶中,大部分酶带活性随染色体倍性增加而减弱,表明基因剂量与POD同工酶酶谱的表达呈负相关.     

红花的愈伤组织诱导及其与过氧化物酶和酯酶同工酶的关系

红花外植体培养于附加不同激素的 MS培养基中,根、胚轴、子叶均可大量形成愈伤组织和根。5个红花品种间过氧化物酶和酯酶同工酶酶谱的差异小于不同器官间的差异。随外植体子叶脱分化过程的进行,过氧化物酶同工酶酶带逐渐增多,而酯酶同工酶酶带则逐渐减少,并出现特异的诱导酶带。     

芒果属植物过氧化物酶同工酶的研究

采用聚丙烯酰胺凝胶电泳法,测定了芒果属１７个种群近缘植物过氧化物酶同工酶,并根据酶谱分布及酶谱特征,结合不同的表现形式,分析了属内种间的亲缘关系。结果表明,异（同）地种间、不同器官酶谱数目、酶含量、迁移率（Ｒｆ）、酶活性及分布特征均有不同程度的差异,可以作为芒果属种间亲缘关系的生化遗传指标。     

芒果生长发育过程中若干生理参数的变化

研究结果表明：芒果植（苗）株过氧化物酶同工酶基本酶谱共不１０条,酶带面度２．６７ｃｍ,同一器官生育期不同,酶谱、Ｒｆ值、酶活性和酶带分布均有明显差异;果实不同成熟阶段,呼吸跃变与乙烯产生相关密切。     

张杂谷苹果酸脱氢酶和过氧化物酶同工酶研究

以张杂谷‘3号’、‘5号’、‘6号’及其亲本为材料,通过聚丙烯酰胺垂直板凝胶电泳技术研究不同材料不同生育期苹果酸脱氢酶和过氧化物酶同工酶酶谱特性,同时对总酶活进行了测定,探讨谷子杂种优势的形成机理。结果表明:(1)张杂谷及其亲本叶片苹果酸脱氢酶共有5条酶带,有共同酶带和偏母本型酶带;过氧化物酶同工酶共有14条,属于共同酶带;从负极到正极苹果酸脱氢酶同工酶Rf分别为0.611、0.626、0.642、0.684和0.716,而过氧化物酶同工酶Rf分别为0.08、0.21、0.24、0.30、0.36、0.44、0.49、0.58、0.66、0.69、0.72、0.75、0.85和0.89。(2)杂种苹果酸脱氢酶同工酶和母本酶谱同型,过氧化物酶同工酶酶谱和父母本一致;供试品种苹果酸脱氢酶酶谱相对简单,酶带集中,而过氧化物酶同工酶酶带数量、活性和宽度差异性较大,酶谱比较复杂。(3)所有品种苹果酸脱氢酶总活性在抽穗期最高,过氧化物酶总活性在灌浆初期最高。杂种苹果酸脱氢酶活性在苗期和抽穗期表现出不同程度的超亲优势,而过氧化物酶在抽穗和灌浆初期均表现出不同程度的超亲优势,这可能与杂种优势有关。     

Selection and characterization of somatic hybrids between Lycopersicon esculentum and Lycopersicon peruvianum

Somatic hybrids of the cultivated tomato, Lycopersicon esculentum, and a wild species, L. peruvianum, were obtained by fusion of leaf protoplasts from both species in the presence of poly-ethylene-glycol (PEG) or in an electric field. The somatic hybrids were selected on the basis of kanamycin resistance of L. esculentum and the plant regeneration capacity of L. peruvainum. Chromosome counts in root tips and the determination of the number of chloroplasts in guard cell pairs revealed that the majority of these hybrids was tetraploid (2n = 4x = 48). The remaining hybrids were at the hexaploid level with chromosome numbers between 64 and 72. The hybrid nature of the regenerated plants was confirmed by analysis of isozyme markers and by their morphology. Most hybrids did flower and set fruits and seeds after selfing. According to RFLP analysis 6 out of the 10 hexaploid hybrids contained two genomes of L. esculentum and four genomes of L. peruvianum. One of these hexaploids had genomes of two different L. peruvianum genotypes and was therefore considered to be derived from a triple protoplast fusion. The hexaploid plants were less fertile than the tetraploids and more resembled L. peruvianum.     

Relationship between tomato fruit growth and fruit osmotic potential under salinity

To investigate the relationship between fruit growth and fruit osmotic potential (Ψ_s) in salty conditions, a sensitive tomato cultivar (Lycopersicon esculentum Mill.) and a tolerant accession of the wild species Lycopersicon pimpinellifolium Mill. were grown in a greenhouse with 0 and 70 mM NaCl, and the growth of the fruit studied from 15 to 70 days after anthesis (DAA). L. pimpinellifolium did not reduce significantly fruit weight in salty conditions throughout the growth period, whereas L. esculentum fruit weights decreased significantly with salinity from 45 DAA. L. esculentum fruit fresh weight reductions resulted from both less dry matter and water accumulation, although the fruit water content was affected by salinity before the fruit weight. In both species, fruit osmotic potential (Ψ_s) decreased significantly with salinity during the rapid fruit growth phase, although the changes were different. Thus, fruits from L. pimpinellifolium salt treated plants showed a Ψ_s reduction at the beginning (15 DAA) twice as high as that found in L. esculentum. As the advanced growth stage (from 15 to 55 DAA), the Ψ_s reduction percentages induced by salinity were quite similar in L. pimpinellifolium fruits, while increased in L. esculentum. Under saline conditions, the solutes contributing to reduce the fruit Ψ_s during the first 55 DAA were the inorganic solutes in both species, while in the ripe fruits they were hexoses. L. esculentum fruits accumulated K⁺ as the main osmoticum in salty conditions, while L. pimpinellifolium fruits were able to use not only K⁺ but also the Na⁺ provided by the salt.     

Antioxidants and antioxidative enzymes in wild-type and transgenic Lycopersicon genotypes of different chilling tolerance

The Mehler–Ascorbate–Peroxidase cycle is a protection system against reactive oxygen species (ROS) occurring during over-excitation of the photosynthetic apparatus. In the cultivated tomato, Lycopersicon esculentum, long-term chilling under moderate light leads to oxidation of the Calvin cycle key enzyme, ribulose-1,5-bisphosphate carboxylase (rubisco), presumably by generation of ROS. In contrast, high-altitude lines of the wild tomato species L. peruvianum were tolerant against the same chilling stress. In the present study, we analysed leaf contents of antioxidants (ascorbate, glutathione) and activities of enzymes of the Mehler–Ascorbate–Peroxidase cycle in the two Lycopersicon species. While antioxidant levels and activities of chloroplast superoxide dismutase (SOD) and ascorbate peroxidase (APX), both inducible by chilling stress, were similar in chilling-tolerant and chilling-sensitive genotypes, chilled L. esculentum showed lower glutathione reductase (GR) activities than high-altitude L. peruvianum. We constructed transgenic plants overexpressing an Escherichia coli GR in the chloroplast (approximately 60-fold of the wild-type (WT) activity). However, these plants resembled identical chilling sensitivity of the photosynthetic apparatus as WT plants as measured after a photoinhibition treatment and by the effect of long-term chilling on rubisco activity. We conclude that the Mehler–Ascorbate–Peroxidase cycle is not the limiting factor for the sensitivity of the photosynthetic apparatus of L. esculentum towards long-term chilling under moderate light. We suggest that a possible cause for the higher chilling tolerance of L. peruvianum is prevention of ROS formation by better conversion of light energy to photochemistry at suboptimal temperatures.     

The induction of lincomycin resistance in Lycopersicon peruvianum and Lycopersicon esculentum

Lincomycin-resistant calli were induced from both Lycopersicon esculentum and Lycopersicon peruvianum using N-mitroso-N-methylurea (NMU) mutagenesis. From these calli lincomycin-resistant plants were regenerated. For L. peruvianum it was shown that the resistant plants could be divided in two classes with respect to their resistance to lincomycin and its derivative clindamycin. The first class comprised plants which were resistant to 500 mg/l lincomycin and showed no shoot or root formation in the presence of clindamycin; the second class consisted of plants resistant to 2000 mg/l lincomycin and these plants were able to form shoots and roots on clindamycin containing media. Lincomycin is an inhibitor of peptidyltransferase; chloroplast encoded parts of this enzymatic function are sensitive for this antibiotic. Reciprocal crosses between our lincomycin resistant and wild type L. peruvianum plants indicated a maternal inheritance of the mutation.     

Somatic hybridization between Lycopersicon peruvianum and L. pennellii: Regenerating ability and antibiotic resistance as selection systems

Somatic hybrids were obtained between the reproductively-isolated tomato species Lycopersicon peruvianum and L. pennellii. Leaf protoplasts of the former species and protoplasts from cell suspension cultures of the latter were fused with polyethylene glycol. A double selection scheme for fusion products was used on the basis of regeneration ability in L. peruvianum and resistance to the antibiotic G418 (2-deoxystreptamine) in an L. pennellii cell line. One tetraploid and four hexaploid hybrids were obtained from this fusion. The hexaploids might have originated by fusion of two L. pennellii protoplasts and one L. peruvianum protoplast. The hybrids were identified on the basis of isozymes (loci Prx-1, Prx-2, Prx-4, Prx-6, Prx-7, Pgi-1 and putative locus Mdh-1), leaf, flower morphology and epidermal hairs. The expression of antibiotic resistance and regeneration ability in the hybrids indicate that these are dominant or codominant traits. The sterility and subvitality of the resulting hybrids questions the value of somatic hybridization as a useful breeding approach in Lycopersicon.     

Transformation of callus cultures of nine plant species mediated by agrobacterium

A simple method for the Agrobacterium-mediated transformation of callus cultures of nine plant species, Lycopersicum esculentum Mill, Petunia hybrida Vilm, Pimpinella anisum L., Solanum melongena L., S. tuberosum L., Nicotiana glauca Graham, N. glutinosa L., N. plumbaginifolia Viviani and N. tabacum L., is described. Plant calli were resuspended in liquid media, co-cultivated with A. tumefaciens, and plated on restrictive media. The combination of a gene for kanamycin resistance and a gene for firefly luciferase was convenient in the selection and confirmation of hundreds of transformants. Four strains of A. tumefaciens, A208, A348, A281, and PC2760, were employed. All of the callus cultures were successfully transformed with at least one strain of A. tumefaciens, and A281 was the most effective of the four strains. N. glutinosa, N. plumbaginifolia, N. tabacum, P. hybrida and L. esculentum were transformed more efficiently than the other species tested.     

EFFECT OF TEMPERATURE AND DAYLENGTH ON PEROXIDASE AND MALATE (NAD) DEHYDROGENASE ISOZYME COMPOSITION IN TOBACCO LEAF EXTRACTS

Tobacco plants were grown in controlled-environment chambers with night/day temperatures of 10/15 C., 20/25 C., and 30/35 C., and with light durations of 6 hr, 12 hr, and 18 hr. Peroxidases and malate (NAD) dehydrogenases were extracted from green leaf tissue and analyzed for isozyme patterns by disc electrophoresis. A total of 18 anodic peroxidase bands were distinguishable—each alteration in a single environmental variable producing a different isozyme profile. Malate (NAD) dehydrogenase isozyme profiles resulting from each environmental condition exhibited at least four major components, but differences in daylength and temperature conditions changed the relative banding intensities and shifted migration rates of some bands. The physiological implications of these findings are discussed.     

A biochemical approach toward the systematics of the Leptothorax “muscorum”Group in North America (Hymenoptera: Formicidae)

Enzyme patterns have been used to distinguish among a number of morphologically very similar ant species belonging to the Leptothorax “muscoru” group in North America. Not counting the already described species L. retractus, L. sphagnicolus and L. crassipilis, the complex apparently consists of at least three or four more different taxa.     

Rapid identification of Leishmania species from formalin-fixed biopsy samples by polymorphism-specific polymerase chain reaction

The precise identification and classification of Leishmania species is important for public health surveillance since different species cause different clinical features of the disease. A highly specific polymerase chain reaction (PCR) panel was developed to enable the identification of the five major Leishmania species that cause New World cutaneous leishmaniases. The primers used for this panel were designed to distinguish the polymorphism in sequences of commonly amplified DNA bands of the parasites produced by arbitrarily primed PCR. These polymorphism-specific PCR diagnoses were performed with formalin-fixed biopsy specimens of the leishmanial lesions from four patients in Ecuador and one hamster skin lesion, and these lesions were determined to be caused by Leishmania (Viannia) panamensis, L. (Leishmania) mexicana, and L. (L.) amazonensis. The PCR panel may offer an important and practical approach to the standardized identification of Leishmania species in field examinations.     

Isozyme studies in Indian mustard (Brassica juncea L.)

Summary A comparative study of peroxidase and esterase isozymes was carried out at five developmental stages of siliqua in order to characterize twelve genotypes of Indian mustard. The studies showed nearly the same number of isozyme bands at every stage for peroxidase and a varying number of isozyme bands for esterase. The appearance and disappearance of bands, along with their intensity scores, indicated the role of different isozymes at different stages of siliqua development. It has been ascertained that these patterns, especially the intensity scores, can be successfully used to characterize different Indian mustard genotypes.