The Histochemical Localization of Several Enzymes of Soybeans Infected with the Root-knot Nematode Meloidogyne incognita acrita

The sites of activity of alkaline phosphatase, acid phosphatase, esterase, peroxidase, adenosine triphosphatase, and cytochrome oxidase were demonstrated histochemically in fresh sections of ''Lee'' soybeans infected by the root-knot nematode Meloidogyne incognita acrita. Each of the six enzymes was more active at the sites of infection than in adjacent non-infected tissue. During the early stages of infection, an increase in enzyme activity was observed in several cells in the proximity of the lip region of the nematode. However, when definite syncytia were observed, increased enzyme activity was confined primarily within the limits of the syncytium. Increased activity paralleled syncytial development and nematode maturation.     

Histochemical Localization of Enzymes in the Cucurbitaceae Acid Phosphatases: Acid Phosphatases

Histochemical localization of acid phosphatases was successfullycarried out on the steins of Cucumis, Cucwbita and Colocynthis—threeof the most important genera of the Cucurbitaceae whose speciesare cultivated for their fruits in the tropics. The localizationsshowed generally widespread activity and similarity in distributionfor this enzyme in the tissues of these plants. For each plantacid phosphatase activity decreased with age. The possible significanceof these localizations has been discussed in the light of thedistributional evidence and the roles usually ascribed to thisenzyme. Conventional histochemical techniques for the localizationof acid phosphatase have also been evaluated.     

Histochemical Localization of {beta}-Glycerophosphatase Activity in Young Root Tips

Glycerophosphatase activity has been studied in frozen sectionsof maize, barley, and broad-bean root tips by the Gomori leadsulphide precipitation procedure. The enzyme was largely localizedat particulate sites in the cytoplasm, and in the cell walls.In maize and barley the particles were similar to acid phosphatase-containingspherosomes found in other tissues and were most active in thecortical cells. The wall reaction was highest in the outer cells,in agreement with biochemical studies. When excised roots wereincubated in the Gomori medium, staining was restricted to thesurface cells. The possible function of this surface activityand its relevance to ultrastructural studies is discussed.     

Intracellular Localization of Several Enzymes in Candida tropicalis Grown on Different Carbon Sources

Intracellular localization of several enzymes related to tricarboxylic acid cycle was investigated during the aerobic growth of Candida tropicalis on acetate, n-alkane and glucose. NADP-linked isocitrate dehydrogenase in acetate-grown cells was mostly found in S₂ fraction (20,000 × g supernatant fraction of protoplast lysate), whereas more than half of this activity in n-alkane-grown cells was recovered in P₂ fraction (20,000 × g pellet fraction). Large parts of NAD-linked isocitrate dehydrogenase and malate dehydrogenase were present in P₂ fraction, while NADP- and NAD-linked glutamate dehydrogenases were found preferentially in S₂ fraction, irrespective of the growth substrates used. Isocitrate lyase was detected in both fractions. Citrate synthase and aconitase in acetate-grown cells were almost particulate. Catalase activity recovered in P₂ fraction was far higher in alkane-grown cells than in acetate- or glucose-grown cells.     

鳜鱼消化道黏液细胞和6种酶的组织化学定位

采用阿利新蓝-过碘酸雪夫氏(AB-PAS)染色和酶组织化学方法对鳜鱼消化道各部位黏液细胞和6种酶的分布与定位进行了研究。结果显示,黏液细胞可为分为4种类型,食道黏液细胞多数为Ⅲ型和Ⅳ型,未见Ⅰ型和Ⅱ型;胃贲门和胃幽门黏膜上皮仅有Ⅰ型黏液细胞;胃体黏膜上皮则以Ⅲ型细胞为主;幽门盲囊中主要为Ⅱ型细胞;前肠和中肠中Ⅳ型黏液细胞最多,Ⅰ型最少;后肠黏液细胞则以Ⅳ型和Ⅱ型为主。酸性磷酸酶(ACP)主要分布于幽门盲囊和前肠的黏膜上皮;碱性磷酸酶(ALP)主要分布于食道、幽门盲囊和整个肠道黏膜上皮;非特异性酯酶(NSE)主要分布于胃幽门、中肠和后肠黏膜上皮;过氧化物酶(POX)在胃幽门黏膜上皮中活性较高;琥珀酸脱氢酶(SDH)主要分布于胃腺中;腺苷三磷酸酶(ATPase)在消化道各部位均有较多分布。鳜鱼消化道黏液细胞和酶的分布型与其它动物有相似之处,也有其一定的特异性,与消化道不同部位的消化吸收机能相适应。     

华北地区油松种群遗传多样性分析

用酸性聚丙烯酰胺凝胶（APAGE）技术分析了华北地区5个油松种群在醇溶蛋白水平上的遗传多样性。95份材料分离出12条带,其中11条具有多态性,得到23种不同的带型。种群多态位点比例在16.67%～66.67%之间。Shannon信息指数和Nei’s指数的统计结果表明：关帝山油松种群的遗传多样性高于其它种群;华北地区油松种内遗传多样性为0.248 9;种群内的遗传多样性为0.150 7,占总变异的60.55%;种群间的遗传多样性为0.098 2,占总变异的 39.45%。     

黄龙山油松林和油松+辽东栎混交林物种组成及优势种群动态

根据黄龙山油松(Pinus tabulaeformis)林和油松+辽东栎(Quercus liaotungensis)混交林共6个群丛中27个典型样地调查资料,分析了松栎林群落物种组成、多样性指数和优势种群动态.结果表明:(1)松栎混交林中灌木层和草本层植物种数均大于油松林,且灌木和草本种类因群落生境不同而有差异.(2)物种多样性分析显示,油松林和松栎混交林中由于生境不同导致灌木层物种多样性差异最大.(3)优势种群分析表明,油松林中阴坡幼苗数量较多,种群最大的为Ⅳ和Ⅴ龄级个体;松栎混交林中油松和辽东栎幼苗充足,到Ⅱ、Ⅲ龄级二者均大量减少,此后油松个体增加而辽东栎减少,说明松栎林发育过程中各层次植物物种相互制约,优势种群之间也存在竞争,且受生境条件影响较大.(4)主成分分析表明,气温、土壤、光照和总郁闭度是主要影响因子.因此,在未来森林抚育中应适当减少优势种群密度,促进异龄化;采取有效措施合理搭配灌木和草本,强化对森林发育有利的因素,提高林分质量.     

Further Studies on the Lyo and Desmo Components of Several Hydrolytic Enzymes and Their Histochemical Significance

This report describes additional studies of the lyo and desmo components of esterase, alkaline phosphatase, acid phosphatase, leucine aminopeptidase, and β-glucuronidase. The techniques used have already been reported (7). Enzyme diffusion occurs to different degrees in different fixatives, and varies somewhat with each enzyme. Loss of enzymatic activity during fixation occurs as a result of both inactivation due to the chemical reaction of the fixative with the enzymic protein, and diffusion of the lyo component into the fixative. The amount of diffusion into formalin can be reduced by the addition of salts, sucrose, or methocel. The pH of the aqueous medium significantly influences the removal of the lyo fraction from the tissue section. A striking similarity can be noted in the proportions of each fraction of enzyme present in the kidney of the rat, dog, and man. The procedure of fixation and paraffin embedding of tissue blocks does not wholly prevent the diffusion of the lyo component from the tissue sections when they are subsequently immersed in the aqueous incubation medium.     

白皮松成熟胚的离体培养研究

以白皮松成熟胚为外植体诱导再生小植株.试验结果表明,成熟胚不定芽诱导以MS培养基最佳,附加0.294 mg/L的NAA和3.56 mg/L的6-BA时,诱导率接近100%;MS培养基附加NAA(0.05 mg/L)时,平均增殖系数可达6.3以上.不定芽增殖率最大值(10)出现在SH培养基上,此时NAA浓度为0.05 mg/L、6-BA浓度为4 mg/L.MS培养基中加入适量活性炭和GA3能促进不定芽生长,随着活性炭和GA3浓度的增加,有效嫩梢(≥2cm和≥4 cm)的比率显著增高;当活性炭和GA3浓度过高时(分别超过2.75 g/L和4.1 mg/L),不定芽的伸长与生长受到抑制.在离体培养条件下,以种胚为外植体获得了无根苗.     

Localization of Enzymes in Mycoplasma.

Pollack, J. D. (University of Connecticut, Storrs), Shmuel Razin, and Robert C. Cleverdon. Localization of enzymes in Mycoplasma. J. Bacteriol. 90:617-622. 1965.-Cells of eight parasitic and two saprophytic Mycoplasma strains were lysed by use of osmotic shock, and the membranes were separated from the soluble fraction by use of differential centrifugation. Cell fractions were tested for reduced nicotinamide adenine dinucleotide (NADH(2)) oxidase, reduced nicotinamide adenine dinucleotide phosphate (NADPH(2)) oxidase, glucose-6-phosphate dehydrogenase, adenosine triphosphatase, ribonuclease, and deoxyribonuclease activities. Adenosine triphosphatase was confined to the membrane fraction of all Mycoplasma strains. The NADH(2) oxidase activity was associated with the membranes of the saprophytic M. laidlawii and with the soluble fraction of the parasitic Mycoplasma strains. NADPH(2) oxidase activity was detected only in the soluble fraction of the parasitic strains. Glusose-6-phosphate dehydrogenase was demonstrated only in the soluble fraction of M. laidlawii. Ribonuclease activity was found usually in both membrane and soluble fractions, but was generally higher in the membrane fraction. In the human and bovine Mycoplasma strains, deoxyribonuclease activity could not be demonstrated in the soluble fraction; in the remaining strains, activity was highest in the soluble fraction. Dissolution of M. laidlawii strain B membranes by sodium deoxycholate significantly increased membrane-NADH(2) oxidase and adenosine triphosphatase activities.     

Histochemical Localization of Phosphatases in Mycoplasma gallisepticum

Histochemical studies of adenosine triphosphatase and acid phosphatase activity were performed on Mycoplasma gallisepticum. The adenosine triphosphatase activity appears to be localized in the bleb and infrableb regions exclusively and is associated with the cell membrane; acid phosphatase activity is localized in the infrableb region and does not appear to be membrane-associated. These findings are consistent with data from biochemical studies of Mycoplasma cell fractions but, unlike them, reveal that adenosine triphosphatase activity is restricted to a particular part of the cell membrane.     

油松雌球果的发生和发育研究

运用薄切片技术对油松（Ｐｉｎｕｓ ｌａｂｕｌａｅｆｏｒｍｉｓ Ｃａｒｒ．）雌球果的发生和发育进行了研究。结果表明：８月初,雌球果原基发生,共外部形态发生明显变化,但内部细胞组织学分区结构与营养茎端结构相似;１０月中旬,雌球果原基的鳞片叶腋处产生最裨的苞片原基,以后苞片原基由基部向顶端连续发生。此时球果原基的顶端结构姨生变化,顶端分生组织区、中央母细胞区和周围分生组织区衍化为套层,肋状分生组织衍化为     

The Variations of the Starch in Pinus tabulaeformis during Embryogenesis

The presence or absence and the distribution of the starch during embryogenesis of Pinus tabulaeformis have been investigated by using the usual histochemical methods. The embryogenesis of the pine may be divided into six stages. In the course of embryo development the accumulation of starch has been observed in the nueellus, the megaspore mother cell and the megaspore, in the female gametophyte, in the pollen grain and in the embryo itself. The larger starch region, however, is only seen in the free apex of the ovule, the female gametophyte and the root cap-suspensor region of the embryo. During embryogenesis the starch region in the free apex of the nucellus appears before the pollination and persists until the late stage of the development of the young embryo. In the female gametophytc the starch region appears about the time of fertilization or at the early stage of proembryogenesis. It is always surrounding the developing young embryo, moves progressively toward the chalazal end of the female gametophyte after the development of the young embryo and then disappears when the embryo is approaching maturity. The starch region in the embryo itself appears in the basal region of the young embryo before the differentiation of the root initials and then develops into the root cap suspensor starch region and finally disappears until the complete development of the embryo and the seed maturity. The starch region, therefore, is the main site for supplying the carbonhydrate for the developing pine embryo. Many young embryos are distributed in the narrow and long embryonal cavity of the female gametophyte during the stage of cleavage polyembryony. But usually only one of them which can obtain the sufficient saeeharide material becomes the dorminant embryo. It is, therefore, considered that the carbonhydrates may be one of the important factors involving the embryo selection. In the mature seed, with the exception of a few residual starch grains scattered in the root cap-suspensor region, no starch grains are found in other organs or 9issues of the embryo. Hence, we may conclude that the starch grains participate actively in the carbonhydrate metabolism and finally they are almost entirely consumed off and they are, therefore, not the reserve material of the pine seed.     

不同生态条件下油松(Pinus tabulaeformis)菌根根际土壤微生物群落

利用BIOLOG代谢指纹方法分析了陕南商南和陕北安塞不同生态条件下油松菌根根际土壤微生物群落。结果表明,安塞油松和商南油松菌根根际微生物对糖类和氨基酸类碳源较易利用,商南油松菌根根际微生物总体上代谢碳源的种类和活性远大于安塞油松,而且对同类碳源的代谢商南油松的AWCD比安塞油松均高出2倍多。安塞油松菌根根际微生物以氨基酸类代谢群为优势类群,商南油松以糖类代谢群为优势类群。微生物群落多样性指数和微生物群落主成分分析（PCA）指标均表明商南油松和安塞油松菌根根际土壤微生物群落有明显不同,起分异作用的碳源主要为糖类,其次是羧酸类和氨基酸类。商南油松菌根根际土壤微生物群落AWCD极显著高于安塞油松（P〈0．01）,细菌数量显著高于安塞油松（P〈0．05）,Shannon指数和丰富度指数达极显著性差异（P〈0．01）,商南油松和安塞油松菌根侵染率差异不显著,但菌根生物量差异达极显著水平（P〈0．01）。相关性分析表明,菌根生物量与丰富度指数、AWCD呈极显著正相关,与Shannon指数呈显著正相关,但是与菌根侵染率相关性不显著。在商南温暖潮湿丘陵区油松菌根根际微生物活性、群落大小和多样性高于安塞油松,在安塞黄土高原干旱区微生物群落稳定性强于商南油松。     

Paternal cytoplasmic transmission in Chinese pine (Pinus tabulaeformis)

Summary. In this paper, the stages of normal sexual reproduction between pollen tube penetration of the archegonium and early embryo formation in Pinus tabulaeformis are described, emphasizing the transmission of parental cytoplasm, especially the DNA-containing organelles – plastids and mitochondria. The pollen tube growing in the nucellus contained an irregular tube nucleus followed by a pair of sperm cells. The tube cytoplasm contained abundant organelles, including starch-containing plastids and mitochondria. The two sperm cells differed in their volume of cytoplasm. The leading sperm, with more cytoplasm, contained abundant plastids and mitochondria, while the trailing one, with a thin layer of cytoplasm, had very few organelles. The mature egg cell contained a great number of mitochondria, whereas it lacked normal plastids. At fertilization, the pollen tube penetrated into the egg cell at the micropylar end and released all of its contents, including the two sperms. One of the sperm nuclei fused with the egg nucleus, whereas the other one was retained by the receptive vacuole. Very few plastids and mitochondria of male origin were observed around the fusing sperm and egg nuclei, while the retained sperm nucleus was surrounded by a large amount of male cytoplasm. The discharged tube cytoplasm occupied a large micropylar area in the egg cell. In the free nuclear proembryo, organelles of maternal and paternal origins intermingled in the neocytoplasm around the free nuclei. Most of the mitochondria had the same features as those of the egg cell, but some appeared to be from sperm cells and tube cytoplasm. Plastids were obviously of male origin, with an appearance similar to those of the sperm or tube cells. After cellularization of the proembryo, maternal mitochondria became more abundant than the paternal ones and the plastids enlarged and began to accumulate starch. The results reveal the cytological mechanism for paternal inheritance of plastids and biparental inheritance of mitochondria in Chinese pine. Correspondence and reprints: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Science, China Agricultural University, Beijing 100094, People’s Republic of China.     

北京地区油松胚珠发育时期的确定

采用显微观察法对北京地区不同发育时期的油松胚珠进行解剖结构观察,以确定北京地区油松胚珠的发育时期。结果表明,北京地区油松胚珠的发育历时13个月(从2003年4月~2004年5月),具体时期为:2003年4月12日为胚珠分化期,4月21日左右为大孢子母细胞时期,5月至8月为游离核时期,2003年8月至2004年2月为胚珠休眠期,2004年4月15日左右进入雌配子体细胞化时期,4月21日左右为颈卵器原始细胞活动期,5月8日左右为中央细胞分裂期,5月14日左右为成熟颈卵器时期。本研究结果中油松胚珠经历的发育阶段与前人的研究基本一致,但在时间上存在差异,而且北京地区比辽宁兴城地区油松胚珠发育时期平均提前了大约20~30 d。     

油松优树选择方法的研究

论述了优树选择应遵循优良种源区选择优良林分,优良林分中的选择优良单株的系统选择原则。提出了标准差法选择方法和标准,要求优树胸径大于林分平均胸径加２倍标准差,且优树年平均出长量：胸径大于０．９ｃｍ、树高大于０．５ｍ。根据１９株人工林优树周转的标准地每木实测数据,对小标准地法和大树选优法作了比较;根据３２株优树的选优结果,比较了１－５株大树法选优法之间的相关关系,大树法间相关都显著,但以３株法为最好;