新疆野核桃种质对低温胁迫的生理响应

以新疆巩留野核桃林37份新疆野核桃种质资源的1年生枝条为材料,6份核桃楸为对照,利用人工气候箱模拟春季低温,分别于-20 ℃和4 ℃处理12 h,测定其相对电导率、游离脯氨酸、可溶性糖、丙二醛含量、过氧化物酶活性等生理生化指标,并进行相关分析。采用隶属函数和主成分分析综合评价核桃种质对低温的生理响应。结果表明: 低温胁迫下,新疆野核桃相对电导率、游离脯氨酸含量、可溶性糖含量、丙二醛含量和过氧化物酶活性均呈上升趋势。综合评价其低温抗性与生境相关:巩留野核桃林中沟>东沟>西沟>主沟。新疆野核桃表现出比核桃楸更为耐低温的特性。本研究从37份新疆野核桃种质中筛选出7份耐低温种质,为改良核桃品种和提高核桃耐倒春寒等生长期的突然天气变化的能力提供科学参考。     

低温逆境对不同核桃品种抗氧化系统及超微结构的影响

为揭示核桃抗寒机理,确定核桃抗寒性鉴定适宜的生化指标,以展叶期抗寒性不同的哈特雷、晋龙1号和晋龙2号3个品种1年生枝条的叶片为材料,测定了1℃低温下抗氧化酶活性及超氧阴离子(O2(-))含量的变化,并采用透射电子显微镜观察低温逆境对抗寒性差异大的哈特雷和晋龙2号叶肉细胞超微结构的影响.结果表明:低温胁迫前后抗寒性强的哈特雷叶片中超氧化物歧化酶(SOD)和过氧化物酶(POD)的活性最高,超氧阴离子含量最低,叶肉细胞超微结构较稳定,叶片没有明显冷害症状.抗寒性差的晋龙2号随着低温胁迫时间的延长,3种抗氧化酶活性的下降幅度最大,O2(-)含量始终处于高水平;胁迫72 h时细胞叶绿体普遍膨胀,基粒片层变薄,数目减少,部分叶绿体被膜及质膜清晰度下降,部分顶端小叶叶缘呈水浸状,表现出冷害症状.可见,低温逆境下核桃叶肉细胞超微结构的稳定性与其品种的抗寒性密切相关.SOD、POD活性以及O2(-)含量可作为展叶期核桃抗寒性鉴定的生化指标;低温胁迫下核桃叶片细胞内膜系统的损伤与活性氧积累之间可能存在一定的相互关系.     

核桃属部分种的小孢子发生及核型研究

本试验采用常规压片法,观察了核桃属(Juglans L.)四个种花粉母细胞(PMC)的减数分裂过程和花粉形态,检测了两个种的花粉生活力,分析研究了七个种的核型。结果表明,普通核桃(J.regia)核桃楸(J.mandskurica Maxim.)和黑核桃(J.nigra L.)的PMC减数分裂基本正常,但河北核桃PMC的减数分裂过程都极不正常,供试的七个种,除普通核桃为2C核型外,其余均为2B核型,仅黑核桃一种带有随体染色体。根据核型特点把七个种分成三组,并探讨了组间与组内种间的亲缘演化关系。作者认为,河北核桃应为一种独立的种,核桃科在系统发育上可能来源于染色体基数为8的两群)不同植物。     

早实核桃脂肪积累与酶活性动态及其相关性

以绿岭和绿早2个早实核桃品种为试材,对开花50 d后不同发育时期核仁的脂肪含量与3种相关的酶活性动态进行研究,分析不同发育时期影响核桃脂肪含量的关键酶.结果表明: 2个品种的核仁脂肪积累动态一致,核仁在开花后50 d开始固化,花后60～90 d核仁脂肪含量迅速增加,花后90～120 d增长幅度变缓,花后120～130 d脂肪含量停止增长.利用Logistic模型对核桃脂肪积累进程进行拟合(P<0.01),绿岭脂肪积累盛期为开花后57.8～85.8 d,绿早为开花后67.4～92.1 d.乙酰辅酶A羧化酶(ACCase)、6-磷酸葡萄糖脱氢酶(G6PDH)和丙酮酸激酶(PK)活性均在花后50～100 d呈上升趋势,随后酶活性呈下降趋势.核仁脂肪含量与ACCase活性呈显著正相关;脂肪积累速率与PK活性呈显著正相关;不同发育时期脂肪含量与酶活性的相关性不同.花后50～100 d是核桃核仁脂肪合成旺盛的时期,此时加强田间栽培管理可以提高脂肪含量.在核桃脂肪合成前期G6PDH是影响脂肪含量的主要酶,PK活性影响丙酮酸形成,从而间接影响脂肪的合成.ACCase活性影响了最终的脂肪含量,并在脂肪合成的各个时期均起到重要的调节作用,是影响核桃脂肪合成的关键酶.     

胃蛋白酶和木瓜蛋白酶水解核桃蛋白工艺研究

以核桃粕为原料,以水解度为考察指标,研究胃蛋白酶和木瓜蛋白酶对核桃蛋白的水解作用.结果表明：木瓜蛋白酶为酶解核桃蛋白最佳酶,其最佳酶解条件为底物浓度4％、酶质量分数5％、酶解 pH 值为7．0、酶解温度50℃、酶解时间4 h;在该条件下,木瓜蛋白酶酶解核桃蛋白水解度可高于25．76％.     

核桃凋落叶分解对莴笋抗氧化系统及光合特性的影响

为探讨核桃对农作物的化感作用,该试验采用盆栽法,设置4个凋落叶施用量水平(0、30、60、90g/盆),研究了核桃凋落叶在土壤中自然分解过程中对莴笋(播种后80、100、120和140d)抗氧化系统、光合生理特征及其生长的影响。结果显示:(1)核桃凋落叶处理的莴笋叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性在播种80d时得到促进,在100d时受到抑制,而在120d之后基本恢复至正常水平,并以SOD表现最为敏感。(2)核桃凋落叶处理100和120d时,莴笋叶片可溶性蛋白(SP)含量显著降低,而可溶性糖(SS)含量显著增加。(3)核桃凋落叶处理100、120d时,莴笋叶片净光合速率(Pn)受到显著抑制,各处理气孔导度(Gs)和蒸腾速率(Tr)显著低于对照。(4)核桃凋落叶处理的莴笋株高、地上部分生物量及地上部分占总生物量比重在处理120d时均显著低于对照,在140d时基本恢复正常。研究表明,核桃凋落叶在土壤中分解对莴笋产生的化感作用强度随分解时间延长呈现出逐渐增强后再减弱的变化趋势;莴笋可以通过调控自身的保护酶活性和渗透调节物质含量在一定程度上缓解化感物质伤害,对核桃凋落叶的化感作用有较强的耐受能力,生产中可以在核桃林下进行间作或者套作莴笋。     

核桃离体胚的植株再生

在选育核桃优良单株时一般利用实生苗繁殖,由于自然杂交,常易丧失优良品种的性状。核桃扦插不易生根,嫁接繁殖因接口处易发生氧化褐变,因而成活率较低。为了利用组织培养技术进行核桃的快速无性繁殖,我们(1984)曾从核桃的不同     

核桃青皮分解对小白菜生长及生理特性的影响

采用盆栽试验,研究了不同剂量核桃(Juglans regia L.)青皮在土壤中分解过程中对受体植物小白菜(Brassica rapa L.var.chinensis)幼苗生长及其生理特性的影响,探讨核桃青皮的化感作用机理。结果表明:(1)核桃青皮在施入土壤的75d内,显著抑制了小白菜地上部分生物量的积累。(2)各剂量青皮处理小白菜叶片的叶绿素含量在施入20d时均较CK降低,且随着处理时间延长总体呈减少趋势。(3)各剂量青皮处理小白菜叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)活性在施入20d和75d时大多比对照显著升高,且有随剂量增加而增强的趋势,但过氧化氢酶(CAT)活性则随添加量增大或处理时间延长多无显著变化。(4)小白菜叶片可溶性糖(SS)含量在施入20d时随青皮添加量的增加而增加,后期则表现为显著下降,而可溶性蛋白(SP)和脯氨酸(Pro)含量在整个处理期则呈减少趋势。研究认为,核桃青皮在土壤中分解可能对小白菜的生长和抗性生理指标产生了明显的化感作用,其化感作用强度随其分解时间延长呈先强后弱的变化趋势,且高添加量产生的效应比低添加量快而且强。     

60Co γ射线辐照对鲜食核桃萌芽及相关生理指标的影响

以‘辽河4号'脱皮鲜食核桃果实为试材,经60Co γ射线辐照处理和聚乙烯(PE)保鲜袋包装后贮藏于(25±3)℃和(20±3)℃条件下,测定了相关生理指标的变化,探究60Co γ射线辐照对鲜食核桃萌芽生理的影响.结果表明,在(25±3)℃和(20±3)℃的贮藏条件下,对照组鲜食核桃分别在贮藏12和20 d 时开始发芽,处理组核桃在贮藏期间未出现发芽现象;在对照组鲜食核桃胚芽萌发期间,其呼吸强度、可溶性蛋白质和游离氨基酸含量及脂氧合酶(LOX)、过氧化氢酶(CAT)、过氧化物酶(POD)活性总体呈上升趋势,脂肪含量和脂肪酶(LPS)活性则降低; 而60Co γ射线辐照有效地减缓了鲜食核桃脂肪、可溶性蛋白质及游离氨基酸含量的下降速度及其呼吸代谢强度,提高了POD活性,却抑制了LOX、LPS、超氧化物歧化酶(SOD)和CAT的活性,从而阻碍了其胚芽的萌发,以达到延长其贮藏期的目的.     

核桃炭疽菌携带病毒种类鉴定及多样性分析

炭疽菌是核桃主要病害核桃炭疽病的病原,目前对核桃炭疽菌携带的病毒种类及病毒基因组序列信息了解较少。利用宏病毒组测序技术,对分离自我国3个不同省份的25株核桃炭疽菌所携带的病毒基因序列进行发掘、鉴定和分类。经同源比对分析,获得22种病毒基因组序列,其中19种为新病毒。在22种病毒中,有21种为正单链RNA病毒,1种为dsRNA病毒。正单链RNA分别隶属于裸露病毒科（Narnaviridae）、线粒体病毒科（Mitoviridae）和葡萄孢欧尔密病毒科（Botourmiaviridae）,而dsRNA属于Alternaviridae病毒科。RT-PCR验证结果表明22种病毒都能在核桃炭疽菌株中被检测到,且25株炭疽菌的病毒携带率为100%,每个菌株都至少被1-11种病毒侵染。研究结果丰富了炭疽菌属所携带的病毒基因组信息,为后续深入分析炭疽菌真菌病毒的多样性和分子特性奠定基础。     

The Subcellular Localization of Isoperoxidases in Capsicum annuum Leaves and their Different Expression in Vegetative and Flowered Plants

The subcellular localization of leaf peroxidases (EC 1.11.1.7)and their expression in vegetative and flowered plants has beenstudied in Capsicum annuum (var. annuum) in order to assesswhether the expression of new peroxidase isoenzymes can characterizethe floral state which determines the beginning of reproductivedevelopment. The results showed that floral development is accompaniedby a significant increase in the level of soluble (non-sedimentable)leaf peroxidase, independently of leaf position along the internodes,and therefore independently of the leaf age. An analysis ofthe leaf peroxidase isoenzyme patterns along the internodesfor vegetative and flowered plants shows that the increase inperoxidase activity is due to a general increase in the activityof all the pre-existing peroxidase isoenzymes, although isoenzymeB₂ and, especially, isoenzyme A₁ showed a distinctive and majorincrease in activity. These two isoenzymes are mainly ionically-boundto cell walls, probably in equilibrium with the same isoenzymesmoving freely in the cell-wall free spaces. The differs fromother peroxidase isoenzymes, such as isoperoxidase B₆, whichis mainly located in the covalently-bound cell-wall fractionand in mesophyll vacuoles. These results are discussed in thelight of a possible role of cell wall peroxidases as markersof the floral state in Capsicum annuum morphogenesis.Copyright1993, 1999 Academic Press Capsicum, floral state, leaf peroxidases, subcellular localization, vegetative state     

Expression of cucumber stress-related anionic peroxidases during flower development or a cryptic infective process

The striking diversity in the expression pattern of the stress-related anionic peroxidase was observed during development of female cucumber flower. While the isoenzyme Prx3 was accumulated constitutively in the course of flower development, the expression patterns of other two isoenzymes (Prx1 and Prx2) were restricted to the period after flower opening. The virus infection was simulated by careful opening of the intact female flower buds 3 d before anthesis followed by exposition to the glasshouse environment for 3 d. The results obtained in this experiment revealed a marked accumulation of the isoenzyme Prx1 and Prx2 at anthesis. Under normal flower development, the pistils did not accumulate these isoenzymes at this stage. In contrast, the pattern of expression of Prx3 as well as of the pistil-specific peroxidase isoenzyme remained unchanged, confirming a constitutive type of expression. Beside the pistil, a 3-d exposition of the stripped flowers resulted in a marked accumulation of Prx1 and Prx2 isoenzymes also in both adjacent flower organs - the ovary and the pedicel. At the same time of the normal development of female flower these organs did not accumulate these isoenzymes.     

Localization and General Properties of Developing Peach Seed Coat and Endosperm Peroxidase Isoenzymes

Changes of soluble and ionically bound peroxidase and indoleacetic acid (IAA) oxidase activities were followed during peach seed development. Soluble peroxidase activity was located mainly in the embryo plus endosperm tissue, whereas wall ionically bound activities were found predominantly in the integument tissue. The different peroxidase isoenzymes present in the extracts were characterized by polyacrylamide gel electrophoresis and isoelectric focusing; the main soluble isoenzyme of embryo plus endosperm tissue was an anionic isoperoxidase of R _F 0.07. Basic ionically bound isoenzymes were located only in the integument tissue, but two soluble anionic isoenzymes of R _F 0.23 and 0.51 were also present in this tissue. In parallel, peroxidase protein content was estimated specifically using polyclonal antibodies. The kinetic data and the changes of seed IAA oxidase activity during fruit development suggested that basic peroxidase isoenzymes from ionically bound extracts of integument might be involved in IAA degradation. Received September 11, 1997; accepted October 21, 1997     

Effect of UV-C on peroxidase isoenzymes in axillary bud cultures ofVitis species differing in fungal resistance toPlasmopara viticola

The effect of shortwave (250 nm) UV radiation (UV-C) on the level of peroxidase activity and peroxidase isoenzyme patterns in leaves of resistant ([Vitis vinifera x Viris riparia] x Vitis rupestris andVitis rupestris) and susceptible (Vitis vinifera) grapevine species toPlasmopara viticola (downy mildew) was studied. The results show that although UV-C did not produce significant changes in peroxidase activity in susceptible species, and only minor changes in resistant species, treatment with UV-light induces an acidic isoperoxidase (isoperoxidase A₁), capable of oxidising 4-hydroxystilbenes in resistant species. It was named HSPrx 2. Since peroxidase is apparently the enzyme responsible for ε-viniferin synthesis from resveratrol in grapevines, a close relationship between this peroxidase isoenzyme and ε-viniferin synthesis which occurs in grapevine leaves after UV-C treatment must be expected.     

Pollen from cold-treated Nicotiana tabacum buds: Embryogenic capacity,peroxidase activity and partitioning in aqueous two-phase systems

Pollen isolated from fresh and cold treated tobacco (Nicotiana tabacum L. ew. Wisconsin 38) flower buds were separated using aqueous polymer two-phase partitioning and analysed with respect to embryogenic capacity, peroxidase activity and isoperoxidase pattern. Pollen with embryogenic capacity from cold-treated flower buds were enriched in fractions with higher partitioning than those from fresh flower buds, but the amounts were the same. Cold led to a general increase in specific peroxidase activity in pollen fractions enriched in embryogenic pollen, and also to specific changes in the isoperoxidase pattern. Neutral peroxidase species (pI around 7) and alkaline species (pIs around 9) could be related to pollen fractions enriched in embryogenic pollen. The data agree with earlier data showing that the amount of pollen with the potential to form embryos is established at an early stage in flower development, whereas if they really do so depends on how they are pretreated, e.g. by cold treatments of the buds. The latter is also reflected by quantitative and qualitative differences in peroxidase.     

Plant Parenthood in Milkweeds: A Direct Test of the Pollen Donation Hypothesis

Abstract For hermaphroditic plant species whose fruit production is limited by maternal resources, the "pollen donation hypothesis" views large floral displays as an adaptation to enhance the probability of fathering seeds on other plants. This hypothesis has frequently been used to describe the evolution of large floral displays in milkweeds ( Asclepias ). Most tests of the pollen donation hypothesis, however, have used indirect measures, such as flower production or pollen removal, to estimate male reproductive success. To test the pollen donation hypothesis directly, we performed a paternity analysis and determined the number of seeds sired by individual genotypes in a natural population of poke milkweed, A. exaltata , in southwestern Virginia. Seeds sired (male success) and seeds produced (female success) were significantly correlated with flower number per plant (for male success: r = 0.32, P < 0.05; for female success: r = 0.66, P < 0.0001). Functional gender of plants that reproduced both as males and females (N = 17) was not correlated with flower number per plant ( r = 0.35, P>0.05), indicating that plants with large floral displays did not reproduce primarily as males. Percent fruit-set and seed number per fruit were higher in 1986, when levels of pollinarium removal also were higher. Furthermore, several umbels that experienced high pollinator activity selectively matured fruits that contained many seeds. We argue that the evolution of large floral displays in milkweeds is the result of selection to increase overall reproductive success rather than male reproductive success alone.     

Hormonal control of floral abscission in zucchini squash (<Emphasis Type="Italic">Cucurbita pepo</Emphasis>)

In the zucchini squash, Cucurbita pepo, a well coordinated abscission of the female flower during fruit set is essential to obtain a fruit of commercial value. In Spain zucchini is mainly produced in greenhouses in Almería, where high temperatures during the spring-summer period provoke a cultivar-dependent defect in fruits known as the “sticky flower” syndrome. This disorder is characterised by an arrest in growth and maturation of floral organs, and a lack of female floral abscission, thus diminishing fruit shelf-life, commercial quality and value. The aim of the present work was to improve knowledge of the abscission process in C. pepo to better understand the fundamental causes of this disorder. The anatomical analysis of abscission shows a well defined male floral abscission zone (AZ), few hours after anthesis, which differs from the female zone which is not differentiated from the adjacent tissue until the abscission process has begun, and which occurs as a consequence of AZ cell enlargement and the dissolution of their cell walls. To evaluate the role of ethylene and auxins in the regulation of floral abscission in zucchini we performed several treatments, with: ethylene, added as 0.25% ethrel solution; AVG, the inhibitor of ethylene synthesis, at 100 μM; indol-3-acetic acid, 100 μM; and TIBA, the inhibitor of auxin polar transport, at 10 mM. These treatments show that ethylene is an accelerator of zucchini floral abscission, and also promotes abscission in isolated AZs of sticky flowers. On the other hand, IAA delays abscission of the female flowers, whilst the inhibitor of auxin polar transport promotes it. The activity of the cell wall hydrolytic enzymes, polygalacturonase and cellulase, sharply increased just before the shedding of zucchini floral organs (72 h after anthesis). Moreover, both enzyme activities were induced by ethylene, which partly explains the ethylene promoting effect.     

Physiological, biochemical, and molecular characterization of a new female sterile mutant in turnip

A female sterile plant with aborted pistils was previously obtained in an advanced self-pollinated turnip inbred line. Cross progeny with normal wild-type (WT) plants successfully characterized the mechanism of pistil abortion, and the mutant was designated ??tpa?? (turnip pistil abortion). Further morphological investigations showed that the tpa mutant included two phenotypes, pistil abortion partially (PAP) and pistil abortion completely (PAC), and our recent result showed that the tpa trait was controlled by two recessive genes (Wu et al. 2011). In this study, comparisons of the floral organs indicated that corolla size and petal, sepal, filament, and anther length measurements did not significantly differ between the tpa and WT plants. Subsequently, the activities of enzymes (catalase, ascorbate peroxidase, and guaiacol peroxidase) related to the metabolism of reactive oxygen, the O ₂ ^? production rate, as well as the contents of hydrogen peroxide and malondialdehyde were tested and found to be generally higher in tpa floral organs than in the WT. The reverse was true for the flower stems, however. Quantitative differences in peroxidase, cytochrome oxidase, and esterase isoenzyme patterns were also detected among these different organs. These findings suggest that pistil abortion is probably correlated with the unbalanced homeostasis of ROS antioxidant enzymes. Moreover, a specific band was recognized exclusively in the tpa plants using random amplified polymorphic DNA analysis. Sequencing and blasting results indicated that the 878-bp DNA fragment was located in the D1/D2/D3 region, near the 5?? end of the 26S rDNA. This study valuably adds to the current knowledge of and will help further elucidate the biological mechanism of female sterility induction.     

Variations of flower size and reproductive traits in self-incompatible <Emphasis Type="Italic">Trollius ranunculoides</Emphasis> (Ranunculaceae) among local habitats at Alpine Meadow

For plants that rely on animals for pollination, the ability to attract the animals to their flowers can be a crucial component of fitness. A large number of studies have documented pollinators to be important selective agents driving the evolution of flower size and correlated traits on a large scale. In this paper, we studied variations of reproductive traits in self-incompatible Trollius ranunculoides (Ranunculaceae) among local habitats at Alpine Meadow. The results showed significant variations of floral size, seed mass per fruit and sex allocation (male/female mass ratio) between different habitats, where floral size and seed mass was not explained fully by variation of plant size among habitats. It suggested that other factors unrelated to plant size might also influence floral variation. However, in our manipulated experiment, it showed no effects of manipulated floral size not only on visit rate of effective pollinators (bees and flies) but also on female success (seed set, seed mass per fruit), irrespective of flower density. Consequently, we could not conclude that the variation of floral size in T. ranunculoides was due to phenotypic plasticity, or natural selection. But if selection occurred, it should not be mediated by pollinators. It was likely that variation of sex allocation between habitats lead to changes of flower or corolla size, because plant invested much less to male function (female-biased sex allocation and larger single seed mass) in shade habitat (bottom of bush) than other exposed habitats, to gain higher fitness. In addition, high-floral density in T. ranunculoides had a negative effect on service of main pollinator (bees) and female success. This situation would influence the strength of selection on floral size.