深山含笑传粉生物学研究

为明确深山含笑(Michelia maudiae)的传粉生物学特性,该文以西华师范大学校园内的深山含笑为研究材料,采取野外观察法记录了深山含笑的开花动态、访花者及访花频率;用游标卡尺法,测定了深山含笑的异交指数(OCI);用醋酸洋红法,测定了花粉胚珠比(P/O);用套袋和人工授粉法,测定了深山含笑的繁育系统。结果表明:(1)深山含笑为早春开花植物,其种群花期为2~3月,持续30d左右,单花花期5~6d;在单花开放初期,最内层的花瓣未绽开,是适应潮湿或多雨的环境而保护花药与柱头免受雨水的冲刷,提高雌雄性适合度,确保繁殖成功的一种策略;(2)深山含笑的有效传粉者为蜜蜂,访花时间集中在晴朗天气的中午;(3)深山含笑的异交指数(OCI)等于5,花粉胚珠比(P/O)为2 933±50;(4)深山含笑为雄蕊先熟,柱头可授性在开花第2~3天达到最高;(5)套袋和人工授粉表明深山含笑为兼性自交和异交授粉,其中异交授粉的座果率和结籽率显著高于自交授粉。早春开花的深山含笑,其传粉者为单一的蜜蜂,繁育系统为异交,部分自交亲和,结果为引种栽培、良种选育等提供依据更好地利用和保护此资源奠定基础。     

6种含笑属植物抗寒性分析与综合评价

以6种含笑属植物为试材,进行田间冻害调查,拟合Logistic方程计算其低温半致死温度,采用石蜡切片技术,测定9项叶片解剖结构指标,运用隶属函数法对各树种的抗寒性进行综合分析与评价.结果表明: 6种含笑属植物叶片的相对电导率与0～25 ℃低温处理3 h的半致死温度之间呈显著正相关.6种含笑属植物低温半致死温度在-20.48～-8.67 ℃,高低顺序为深山含笑>红花深山含笑>峨眉含笑>杂交11-8>阔瓣含笑>六瓣含笑;叶片表皮细胞1～2层,垂周壁略呈波浪状,栅栏组织细胞1～3层,9项叶片解剖结构指标差异显著,栅栏组织厚度、栅海比和主脉厚度是影响抗寒性的主要叶片解剖结构指标.电导法和隶属函数法对6种含笑属植物的抗寒性评价结果与田间调查结果基本一致.6种含笑属植物抗寒性强弱顺序为:六瓣含笑>阔瓣含笑>杂交11-8>峨眉含笑>红花深山含笑>深山含笑.     

深山含笑叶过氧化物酶对双酚A清除效应研究

深山含笑是一种常绿乔木,具有较好的园林观赏价值,其叶片在每年春天新旧叶交替时,会陆续凋落而不能实现其经济价值;我们前期研究发现深山含笑成熟叶片具有较高的过氧化物酶活性,而过氧化物酶在内分泌干扰物生物酶法降解途径中发挥着重要作用。为充分开发深山含笑叶的经济价值,提高资源的利用效率,我们对深山含笑叶过氧化物酶进行了纯化并初步研究了其性质;在此基础上,进一步研究了深山含笑叶过氧化物酶在降解环境污染物双酚A上的特性。结果显示,经过非离子表面活性剂(吐温-80)抽提、双水相萃取、DEAE-Sepharose离子交换层析,首次从深山含笑叶片中快速分离提纯结合态过氧化物酶,相对可溶性过氧化物酶纯化倍数为55. 2倍,回收率为19%。该酶的比活力为18 999 U/mg蛋白,以愈创木酚为反应底物的最佳pH值为4. 5,最适温度为50℃;具有较宽的pH稳定范围,热稳定性较好,60℃以下不易失活。深山含笑叶过氧化物酶对BPA具有良好的清除能力,在pH4～7,温度30～40℃,H2O2/BPA摩尔浓度之比达0. 8条件下,经过3 h,10 000 U/L POD对0. 2 mmol/L BPA清除率达90%以上,深山含笑过氧化物酶在酚类物质生物酶法催化降解途径中具有潜在的重大利用价值。     

中华猕猴桃单芽腹接试验

我国是猕猴桃的原产地,资源丰富,分布辽阔。但目前还未充分利用,绝大部分尚处野生状态。猕猴桃为雌雄异株植物,花前不易区分,用种子繁殖不能保持品种优良性状,且结果晚;扦插生根困难,成活率低;所以嫁接是进行良种繁育,变野生为人工栽培,迅速实现     

不同砧木阔瓣含笑嫁接苗生长生理及亲和性评价

以白玉兰、紫玉兰、乐昌含笑、深山含笑作砧木的阔瓣含笑嫁接苗为材料,通过对嫁接苗生长特性的观测和生理生化指标的动态测定,运用主成分分析和隶属函数法计算亲和性综合指数,评价阔瓣含笑与4种砧木的苗期亲和性。结果表明:以深山含笑和紫玉兰作砧木的嫁接成活率较高(分别为88.33%、83.33%),白玉兰次之(为75%),乐昌含笑较低(为63.3%);砧木对嫁接苗的苗高有较大影响,以深山含笑和紫玉兰的苗高最高,白玉兰次之,乐昌含笑最矮;砧木对嫁接苗叶片数的影响表现为初期存在一定差异,生长后期差异逐渐缩小。在嫁接苗生长过程中,4种砧木嫁接苗的叶绿素含量均呈双峰型变化,类胡萝卜素含量、可溶性蛋白含量、POD活性表现出先上升后下降的趋势,可溶性糖含量总体表现为先下降后上升的趋势。在4种砧木的嫁接苗中,深山含笑、紫玉兰和白玉兰的各项生理指标都相近,而乐昌含笑整个生长期都低于其它3种嫁接苗。从亲和性综合指数来看,深山含笑的亲和性指数最高(为0.518),乐昌含笑的最低(为0.470)。综合分析,认为4种砧木与阔瓣含笑都具有一定的亲和性,其中以深山含笑最好,乐昌含笑最差。     

红花种质资源色价及主要产量性状的聚类分析

以筛选出来的69份无刺红花资源为材料,利用相关性分析及聚类分析对其花瓣色价、单株果球数、单株果球种子数、果球直径、百粒重、单株种子产量、单株花瓣产量7个性状进行评估。结果表明:69份材料间存在较大的差异,对红花种子产量贡献最大的农艺性状是单株果球数;对红花花瓣色价进行分析,将69份红花资源分为3类,色价在6.3~14.1之间,其中色价极高型材料有14份,平均值达到12.38。对7个性状指标进行综合聚类分析,可以将69份红花资源分为5大类群,各类群性状变化丰富,其中组Ⅳ综合表现最好,是选育高色价、高产红花的优良材料。69份红花资源在色价和产量指标等方面表现出多样性,遗传较丰富,但色价高的资源材料相对较少。红花种质资源色价及主要产量性状的综合分析,可为红花资源的有效利用及选育高色价、高产红花新品种奠定基础。     

低温胁迫下6种木兰科植物的生理响应及抗寒相关基因差异表达

分析了6种木兰科植物对低温胁迫的生理响应及耐寒相关调控基因HSP90和WRKY33的差异表达,为木兰科植物抗寒机理的研究和抗寒品种的选育提供理论基础。结果表明,6种木兰科植物的低温LT50在-10.64—-22.06℃,从高到低依次为红花深山含笑、峨眉含笑、杂交含笑、阔瓣含笑、六瓣含笑和乐东拟单性木兰;低温过程中,6种木兰科植物叶片可溶性蛋白(SP)、游离脯氨酸(Pro)含量、超氧化物歧化酶(SOD)和过氧化物歧化酶(POD)活性呈先升高后降低的趋势,可溶性糖(SS)和丙二醛含量(MDA)则不断积累;筛选出REC、MDA、SP、SS和Pro作为6种木兰科植物抗寒性评价的关键指标;聚类分析将6种木兰科植物在抗寒性能上分为强、中、弱三类,分别为乐东拟单性木兰和六瓣含笑,阔瓣含笑、杂交含笑和峨眉含笑,以及红花深山含笑。对HSP90、WRKY33基因的差异表达分析表明,2个基因在6种木兰科植物中的相对表达量呈先升高后降低的趋势,在临近各树种LT 50时,2个基因的表达被强烈抑制且后期表达量不可逆。0℃时,2个基因的表达量差异不显著;-5℃时,2个基因开始被激活,表达量增加;-10℃时,HSP90、WRKY33基因在红花深山含笑叶片中的表达量较-5℃时分别下调了0.76倍和0.68倍,而在其他5个树种中的表达被进一步激活;-15℃时,HSP90和WRKY33基因在抗寒性中等的阔瓣含笑、杂交含笑、峨眉含笑中亦被强烈抑制,较-10℃时分别下调了0.38倍、0.33倍、0.32倍和0.71倍、0.72倍、0.74倍,在抗寒性强的乐东拟单性木兰和六瓣含笑中的表达被进一步激活;-20℃以后,2个基因在6个树种中的表达均被强烈抑制,但在抗寒性最强的乐东拟单性木兰中的表达量仍高于其他5个树种。抗寒基因的激活与表达是影响植物抗寒性的重要因素,抗寒性不同的树种对低温的应答机制明显不同。抗寒性越强的树种越能快速启动低温应答机制,激活抗寒相关基因的表达,进而调整生理生化活动以抵御和适应冷应力。不抗寒树种中抗寒基因的表达则受到抑制,降低了其对低温逆境的耐受能力。     

极小种群广东含笑野外资源现状

为掌握极小种群广东含笑(Michelia guangdongensis)的种质资源现状并评估其濒危等级, 本文对广东含笑原生地——广东石门台国家级自然保护区及其邻近地区进行了系统的野外调查。结果表明: (1)目前发现广东含笑有4个野外分布区, 其中广东石门台国家级自然保护区红珠岩段和英德上天堂为首次报道。(2)船底顶是广东含笑最集中的分布区, 种群数量最多, 约有1,100株; 英德上天堂种群数量最少, 只有20株。(3)依据IUCN全球物种濒危等级评估标准, 广东含笑应当被评为濒危(EN, B1a + B2a)。     

深山含笑二次开合开花动态与雌雄异熟特征

对深山含笑（Michelia maudiae）的花部特征、开花动态、开花物候、花粉活力和柱头可授性进行研究。结果表明：①深山含笑花期为2月下旬至4月中上旬,表现为集中开花模式;②单花开放时间固定,且有二次开合现象,开放过程可分为露白、首次开花期、闭合期和二次展开期共4个时期;③深山含笑在单株水平上为同步集中式开花,单花花期约为7~12 d,单株花期约为20~38 d;④深山含笑柱头在开花前1 d已具有可授性,开放当天可授性最高,随后可授性迅速下降,而雄蕊在开花第2 d开始散粉,花粉活力最高时期为开花第3 d,为（64.6±6.1）%;⑤深山含笑为雌雄异熟且雌蕊先熟,并且雌雄特征有重叠表达阶段,这样不仅可有效减少自交,又可支持自交,足以保证生殖成功。     

深山含笑大、小孢子发生和雌、雄配子体发育研究

采用石蜡切片技术对深山含笑大、小孢子的发生和雌、雄配子体发育进行观察：深山含笑花药4室,花药囊壁由5-7层细胞构成,腺质绒毡层,小孢子胞质分裂为修饰性同时型,四分体有四面体型、对称型,偶有交叉型,成熟花粉为2细胞型;胚珠倒生、双珠被、厚珠心,大孢子四分体直线型排列,合点端为功能大孢子、雌配子体发育方式为蓼型。从雌、雄配子体发育时间的先后来看,深山含笑春季雌、雄配子体能正常发育,雄蕊先熟,雄蕊和花瓣凋谢后雌蕊大孢子母细胞才形成。而秋季开花的深山含笑,花药中小孢子在孢原细胞或初生造孢细胞期停止发育,花粉败育;雌蕊胚珠珠心组织也未见大孢子母细胞发育,开花后雌蕊随花柄凋落。该研究为深山含笑生殖发育和杂交育种积累了资料。     

Feeding in some Australasian Cymatiidae (Gastropoda:Prosobranchia)

Of the four common Australasian cymatiids studied, Cabestana spengleri and Mayena australasia feed almost entirely on simple ascidians. Monoplex australasiae feeds on bivalves as well as on ascidians and Charonia rubicunda feeds on a variety of echinoderms. All species locate their prey by chemoreception of current born prey substances. Charonia rubicunda is a nocturnal predator while the other species feed almost continually.     

Microassay for biological and chemical control of infection of tobacco byPhytophthora parasitica var.nicotianae

We developed a simple, rapid, small-scale assay for infection of tobacco seedlings byPhytophthora parasitica var.nicotianae. One 7-day-old tobacco seedling was placed in each well of a 96-well microtiter plate and inoculated with 500 zoospores ofP. parasitica var.nicotianae. After 72 h all of the inoculated seedlings of the susceptible cultivar, KY14, were infected, and the pathogen had produced sporangia that were visible on the surfaces of the seedlings. Sporangia did not develop on seedlings that were inoculated simultaneously with zoospores and either 1 µg/mL of the chemical fungicide metalaxyl or 5 µL of filtrate of a sporulated culture of the biocontrol agent,Bacillus cereus UW85. Seedlings of tobacco cultivar KY17 were infected byP. parasitica var.nicotianae, although mature plants of this variety are resistant to the pathogen. This microassay may facilitate the rapid screening of potential biological and chemical control agents and may be useful for studying mechanisms of infection and control ofPhytophthora spp. under hydroponic conditions.     

番茄抗青枯病种苗选育初步研究

通过大量收集野生茄科植物作为栽培番茄的嫁接砧木 ,进行嫁接试验并观察嫁接成活率。栽植各砧木苗、番茄实生苗及嫁接成活率较高的嫁接苗 ,观察比较它们在自然条件下青枯病的发病率 (如果自然条件下青枯病发病率低的地块 ,可考虑人工接种病原 )、植株生长情况、产量和品质等 ,筛选出 2种嫁接成活率高 ,抗青枯病能力强 ,并能保持或提高接穗果产量和品质的番茄嫁接砧木。为深入进行番茄抗青枯病种苗选育研究和生产实践奠定了基础。     

Herbivore-Specific,Density-Dependent Induction of Plant Volatiles: Honest or “Cry Wolf” Signals?

Plants release volatile chemicals upon attack by herbivorous arthropods. They do so commonly in a dose-dependent manner: the more herbivores, the more volatiles released. The volatiles attract predatory arthropods and the amount determines the probability of predator response. We show that seedlings of a cabbage variety (Brassica oleracea var. capitata, cv Shikidori) also show such a response to the density of cabbage white (Pieris rapae) larvae and attract more (naive) parasitoids (Cotesia glomerata) when there are more herbivores on the plant. However, when attacked by diamondback moth (Plutella xylostella) larvae, seedlings of the same variety (cv Shikidori) release volatiles, the total amount of which is high and constant and thus independent of caterpillar density, and naive parasitoids (Cotesia vestalis) of diamondback moth larvae fail to discriminate herbivore-rich from herbivore-poor plants. In contrast, seedlings of another cabbage variety of B. oleracea (var. acephala: kale) respond in a dose-dependent manner to the density of diamondback moth larvae and attract more parasitoids when there are more herbivores. Assuming these responses of the cabbage cultivars reflect behaviour of at least some genotypes of wild plants, we provide arguments why the behaviour of kale (B. oleracea var acephala) is best interpreted as an honest signaling strategy and that of cabbage cv Shikidori (B. oleracea var capitata) as a “cry wolf” signaling strategy, implying a conflict of interest between the plant and the enemies of its herbivores: the plant profits from being visited by the herbivore''s enemies, but the latter would be better off by visiting other plants with more herbivores. If so, evolutionary theory on alarm signaling predicts consequences of major interest to students of plant protection, tritrophic systems and communication alike.     

不同苗龄接穗的西瓜嫁接体愈合过程中的3种酶活性变化

用贴接法嫁接西瓜品种早佳(8424)和砧木葫芦品种将军的结果表明:接穗苗龄越大,嫁接成活率越低.多酚氧化酶(PPO)活性嫁接初期较高,以后变化平稳;大苗龄接穗嫁接体的PPO活性较高.嫁接初期过氧化物酶(POD)和苯丙氨酸解氨酶(PAL)活性升高,以后呈下降趋势,木质素合成和维管组织分化阶段再度升高.     

林隙与林下环境对锐齿槲栎和米心水青冈种群更新的影响

锐齿槲栎 (Quercusalienavar.acuteserrata ) 和米心水青冈 (Fagusengleriana) 是广泛分布在我国暖温带和亚热带山地中海拔地段的主要建群种, 对水土保持和维持生态系统稳定具重要意义。该文通过在神农架地区的典型样地调查, 分析了这 2个种群在林隙和林下不同光环境中的更新状况和径级结构 ;利用便携式Licor₆ 4 0 0光合仪测量了 2种苗木的光饱和及光诱导曲线, 探讨了光对这 2个树种更新的影响机理。实验结果表明, 在所调查的林地中, 锐齿槲栎和米心水青冈种群都有充足的幼苗库, 锐齿槲栎幼苗以实生为主, 米心水青冈幼苗以萌生为主 ;从幼苗到幼树的转化过程中, 锐齿槲栎的成活率几乎为零, 而米心水青冈的成活率约为 6 %~ 8% ;在两者的混交林中, 米心水青冈种群呈稳定的金字塔形年龄结构, 而锐齿槲栎则是某一生长阶段的种群占据主导地位。光饱和曲线显示, 对 2年生同生群苗木, 在林窗下, 锐齿槲栎的最大净光合速率 (4.6 1μmol·m-2 ·s-1) 比米心水青冈的 (4.16 μmol·m-2 ·s-1) 要高 ;而在林冠下, 米心水青冈的最大净光合速率 (Pn=3.89μmol·m-2 ·s-1) 比锐齿槲栎的最大净光合速率 (Pn=3.6 8μmol·m-2 ·s-1) 要高。无论在林窗下或林冠下, 米心水青冈幼苗的光饱和点比锐齿槲栎的要低。两者混交林下的光诱导曲线显示, 与米心水青冈幼苗相比, 锐齿槲栎幼苗对光的反应速度较慢, 最大净光合速率也较低, 在林冠下表现出较弱的竞争力。资源分配试验证明, 锐齿槲栎在幼苗阶段以地上部分生长为主, 而米心水青冈倾向于把更多的干物质贮藏在根部, 显示出不同的光利用策略。     

Competitive interactions between Microdochium nivale var. majus, M. nivale var. nivale and Fusarium culmorum in planta and in vitro

Microdochium nivale var. majus and var. nivale are economically important fungal pathogens of cereal seedlings, stem bases and ears, as is the toxigenic species Fusarium culmorum. Competition experiments on seedlings support an earlier report of differential host preference between the varieties of M. nivale on wheat and rye seedlings at 15 degrees C, but showed that it does not extend across a broad range of temperatures. The studies showed that, although interaction is disadvantageous to the less virulent pathogen, it does not confer an advantage to the more virulent pathogen. In mixed inoculum experiments on wheat seedlings at 15 degrees C and 20 degrees C, F. culmorum suppressed the growth of both varieties of M. nivale. However, if M. nivale var. majus became established on the seedlings, it was able to co-suppress colonization of wheat seedlings by F. culmorum. In contrast M. nivale var. nivale did not suppress F. culmorum significantly. The growth of M. nivale var. majus and F. culmorum was also co-suppressed in liquid culture. Significantly, the accumulation of deoxynivalenol mycotoxin was also reduced in the mixed in vitro culture compared with axenic culture of F. culmorum. However, in vitro interaction studies on solidified media were of only limited use in predicting the outcome of competitions in planta.     

Ten polymorphic microsatellite markers in Michelia maudiae (Magnoliaceae)

? Premise of the study: Michelia maudiae is a threatened species in the Magnoliaceae. Microsatellite markers were developed and characterized in M. maudiae for further investigation of its conservation genetics. ? Methods and Results: Microsatellite markers were developed in M. maudiae using the Fast Isolation by AFLP of Sequences Containing Repeats protocol. Ten polymorphic microsatellites were assessed in two populations of M. maudiae. The number of alleles detected per locus varied from 1 to 8. Observed and expected heterozygosities ranged from 0.000 to 0.792 and from 0.000 to 0.826, respectively. Six primer pairs showed transferability in the two related species Michelia foveolata and Michelia chapensis. ? Conclusions: This set of microsatellite markers provides a useful tool for future studies of the conservation genetics of M. maudiae and other congeneric species.