授粉方式对铁皮石斛种质座果及种子萌发的影响

为进行铁皮石斛(Dendrobium officinale)的良种选育和种苗生产,对46个铁皮石斛种质进行人工授粉试验,研究了不同授粉方式的授粉成功率、座果率和种子的萌发能力。结果表明,4种授粉方式的授粉成功率明显不同,不同种质间的授粉成功率可达100%,且座果率普遍较高。只有31.3%的种质可以成功自花授粉,且座果率相差较大。同株异花授粉和种质内异株授粉的成功率和座果率均介于上述二者之间。授粉后所得种子的萌发率和萌发速度在4种授粉方式间也呈现与授粉成功率和座果率一致的趋势。这表明在铁皮石斛种质中存在自交不亲和现象,因此以不同产地来源的铁皮石斛种质为亲本进行人工授粉,可大大提高授粉的成功率和座果率,以及后代种子的萌发能力。     

大花黄牡丹的开花特性与繁育系统

2019-2020年观察河南省栾川县引种栽培的西藏特有植物大花黄牡丹(Paeonia ludlowii)开花过程和访花昆虫,计算花粉-胚珠比(P/O)和杂交指数(OCI),检测花粉活力与柱头可授性并结合人工控制授粉试验,探究其开花特性和繁育系统类型.结果表明:(1)大花黄牡丹花朵昼开夜合,花朵随花冠展开下垂,伴花冠闭合...     

桔梗开花期花粉活力变化对其结实能力的影响

对桔梗花粉离体萌发培养条件进行了筛选,并采用花粉离体萌发法测定了桔梗花粉发育过程中其活力的动态变化,采用人工授粉法对开花后不同时间花粉的授粉结实能力进行了研究.结果显示:(1)桔梗花粉离体萌发的最适温度为30℃,培养1.5 h是花粉管测定的最佳时间.桔梗花蕾期和开花当时花粉离体萌发率和花粉管长度均较低,开花后花粉继续发育,活力逐渐提高,到花后6 h其萌发率和花粉管长度均达到最高,分别为92.2%和602.9 μm,之后活力逐渐下降,花后120 h桔梗花粉的萌发率为63.1%,而花粉管长度只有287.0 μm,仅为花后6 h花粉管长度的47.6%,开花120 h(5 d)后花粉活力下降速度明显加快.(2)桔梗开花后当天花粉人工授粉的结果率、结籽率最高,之后随着花后天数的增加,结果率和结籽率均迅速下降,开花后3 d的花粉结籽率降为0,已完全丧失授粉结实能力.研究表明,在田间条件下,桔梗花粉寿命可保持5 d以上,而授粉结实能力只有2 d左右;花粉管生长缓慢、长度变短可能是导致花后3 d花粉丧失授粉结实能力的主要原因.     

濒危植物安徽羽叶报春两种花型的繁育特性及其适应进化

二型花柱的维持机制和自然选择压力多年来一直是生态学和进化学研究领域的热点之一。通过实验室栽培和野外观察统计相结合的方法,对安徽羽叶报春两种花型(长柱花和短柱花)的形态特征、花粉活力、柱头可授性、花粉/胚珠比、自然授粉及结籽能力、自交亲和性等繁育特性进行了比较研究。结果表明:长、短柱花的花冠直径和裂片宽无明显差异,而花冠筒、雌蕊和雄蕊高、花粉数目及大小、P/O比均有显著差异。在自然条件下,长柱花所接受的总花粉数要明显高于短柱花的总花粉数,但所接受的异型花花粉数和平均每果结籽数两者无显著差异。长柱花和短柱花的花粉和柱头活力相似,均能在较长时间内维持较高活力,仅在开花末期显著下降。两种花型的花在自花授粉、同型异花授粉、异型花授粉条件下均能结籽,但异型花授粉的结籽数均明显高于自花授粉和同型异花授粉结籽数。在长柱花各种授粉方式中,花粉萌发率无明显差异,但异型花花粉管的生长速度明显比同型异花花粉和自花花粉的快,而在短柱花柱头上表现为异型花授粉的萌发率最高,但只要萌发后在花柱中的生长速度无显明差异。此外,综合上述结果,对二种花型花部综合征的维持机制及自然选择压力进行了探讨。     

超低温保存的梅花花药萌发率和授粉后的结实能力

试验比较梅花花粉和花药超低温保存后的花粉萌发率、授粉结实率和座果率的结果表明:(1)超低温保存花药用室温、自来水冲洗和温水浴3种方法化冻后的花粉萌发率差异不显著。(2)花药超低温保存1h后的花粉萌发率与新鲜花粉差异不显著。(3)超低温保存1个月和1年的花粉和花药都有授粉结实能力。     

花冠对紫茉莉繁殖适合度的影响

以授粉后具花闭合特性的紫茉莉为研究材料,通过在不同天气状况下作去花冠和存留花冠处理,观察花冠对花粉活力、柱头可授性及结实率等繁殖特性的影响。结果表明:紫茉莉自然存留花冠比去花冠的花粉活力、柱头可授性、柱头上的花粉数及柱头上花粉萌发率各指标达最高值的时间重叠性更强,并以阴天存留花冠的最强。存留花冠通过这种时间的重叠性来保证花粉在柱头上的萌发,产生了有利的繁殖适合度。越晚去花冠结实率越高,结实率呈现花闭合自然存留花冠>花冠闭合期去花冠>散粉初期去花冠>花冠展开期去花冠的规律。因此,花冠对紫茉莉繁殖适合度具有利影响。紫茉莉花冠的存留提高了繁殖适合度,增强了对环境的适应能力。     

肉苁蓉人工控制离体寄生关键技术研究

以梭梭无菌苗和肉苁蓉萌发种子为材料,对肉苁蓉离体寄生培养基质、离体寄生方式和离体寄生环境条件等进行了研究,以明确肉苁蓉的寄生特性,为肉苁蓉人工繁殖技术体系的建立与优化奠定基础。结果表明:玻璃纤维滤纸是肉苁蓉离体寄生适宜的培养基质;在黑暗、相对湿度80%、温度18℃～30℃的环境中,萌发的肉苁蓉种子与寄主梭梭寄生培养8d后开始建立寄生关系,寄生率达82%;而光照条件下,萌发的肉苁蓉种子与梭梭不能建立寄生关系。     

人工辅助授粉探寻濒危红树植物红榄李种质资源挽救途径

为了探索濒危红树植物红榄李(Lumnitzera littorea (Jack.) Voigt)种质资源挽救途径, 以三亚铁炉港红榄李为授粉材料, 采集三亚铁炉港和陵水新村港红榄李花粉进行人工辅助异花授粉试验, 结合野外观察和种子萌发实验进行验证。结果表明: 三亚铁炉港红榄李的花期较长, 从花芽开始膨大到落花期末, 最短60 天, 最长可持续140 天, 且开花不整齐; 用培养基法对三亚铁炉港和陵水新村港红榄李的花粉生活力进行检测, 均不超过10%; 人工辅助异花授粉后种子的萌发率有显著提高(自然授粉种子萌发率为0.2‰), 但最高也只能达到5.6‰。实验室萌发种子能够长成幼苗, 且野外林下发现一株红榄李幼苗, 这说明通过人工辅助异花授粉对提高红榄李种子繁育能力有一定的积极作用。     

壳斗科三种植物种子大小对昆虫寄生及种子存活率的影响

种子内的寄生昆虫可以严重影响种子的发育、损害种子活力。种子足余策略理论认为大种子有利于抵御和适应昆虫寄生取食,但动物最优觅食理论推测,大种子更易遭受昆虫寄生。为对这两种对立观点进行验证,本实验以青冈、苦槠和麻栎各2个种群的种子为材料,对昆虫寄生与完好种子间的体积和萌发率进行比较,并对寄生种子萌发率与种子体积的关系进行了分析。结果显示:(1)在6个种群的种子中,只有松阳麻栎和青冈种群的寄生种子体积大于完好种子,其余4个种群的寄生种子体积小于完好种子,但这种差异不显著;(2)所有寄生种子的整体萌发率(18%)显著低于完好种子(45.66%)(P<0.001),在不同种群内,寄生种子的萌发率也分别显著低于完好种子。(3)比较同种植物体积差异显著的寄生种子的萌发率发现,大种子总比小种子具有更高的萌发率,但差异不显著;在不同植物的寄生种子间比较时,体积最大的麻栎种子萌发率显著高于体积较小的青冈和苦槠种子。研究结果表明,象虫在种子上产卵时对大种子没有选择偏好,在昆虫寄生取食严重损害种子活力的压力下,大种子比小种子具有更强的耐受力。     

耐热水稻品种“Nagina 22”高温胁迫下的生理响应

为研究耐热水稻品种Nagina 22(N22)开花结实期高温胁迫下的主要生理响应,采用温室盆栽试验,在各供试材料的开花期,利用人工气候室进行高温胁迫处理。研究结果表明:N22在高温胁迫下不改变开花期但表现出日开花量的转峰;与N22相比,水稻感热品种Moroberekan花药开裂显著受阻,柱头上萌发的花粉数显著减少;花粉萌发数与花药开裂状况呈极显著的相关关系,进而表现出主穗结实率与柱头上花粉萌发数呈显著的相关关系;主穗日结实率在高温下呈递减趋势,且Moroberekan较N22的下降速率显著加快,表明耐热品种与高温胁迫对结实率存在累积效应,而且高温胁迫效应发生在开花授粉之前。     

Biosteres longicaudatus: Developmental dependence on host (Anastrepha suspensa) physiology

Larvae of Anastrepha suspensa that were in the first day of the third instar were parasitized by females of the solitary endoparasitoid, Biosteres longicaudatus. At the end of the 6-hr oviposition period, larvae were ligated posterior to the ring gland so that some larvae had parasitoids anterior to the ligature while in others, the parasitoids were in the abdomen, posterior to the ligature. Ninety-two percent of the parasitoids anterior to the ligature hatched to the first through third instars. Parasitoids posterior to the ligature had a 75% egg hatch to the first instar only. No larval molts to the second or subsequent instars occurred in these parasitoids. Upon parabiosis to 3-day-old, unparasitized host pupae, the ligated larvae pupated and 97% of the first-instar parasitoids in these parabiosed larval abdomens molted to the second instar. Newly laid parasitoid eggs transplanted to 3-day-old pupal hosts had less than one-third of the egg hatch of those transplanted to first-day third-instar hosts. The data implicate the physiological state of the host (vis-a-vis pupation and associated events) as being an important factor in the development of the endoparasitoid.     

Eimeria nieschulzi and Trichinella spiralis: Analysis of concurrent infection in the rat

The effects of concurrent primary infection of the rat with Eimeria nieschulzi and Trichinella spiralis on the number of oocysts of E. nieschulzi shed by the host and on the number, distribution, and fecundity of adult T. spiralis were analyzed. When rats were initially infected with E. nieschulzi followed 9 days later by infection with T. spiralis there occurred a significant decrease in the total numbers of adult worms in the small intestine, a significant shift in the position of these worms along the length of the small gut, a decrease in the fecundity of adult female worms, and a decrease in muscle parasitism when compared with rats infected with T. spiralis alone. When rats were initially infected with T. spiralis, followed 9 days later by infection with E. nieschulzi, there occurred a significant decrease in the numbers of oocysts shed over 24 hr on Days 7, 9, and 11 postinfection below that seen with rats infected only with Eimeria. These changes are discussed in terms of the enteropathophysiologic lesions and enteric inflammation known to occur during infections with these two parasites.     

Tritrichomonas foetus: Stable anaerobic resistance to metronidazole in vitro

Stable anaerobic resistance of Tritrichomonas foetus to metronidazole was induced in vitro by cultivation of trichomonads in the Diamond's TYM medium with metronidazole in concentrations sublethal to the parasites. Nine metronidazole-resistant strains were derived from four drug-susceptible clones of the T. foetus strain KV-1. Subculturing the parasites at both increasing and constant pressure of the drug resulted in development of resistance if the medium contained at least 3 μg ml^?1 of metronidazole and the organisms were exposed to the drug for 3 to 8 months. The development of resistance was gradual and in all clones investigated proceeded through similar sequence of stages: (1) Survival without growth and subsequent reproduction at low metronidazole concentrations (1 to 5 μg ml^?1. (2) Survival and reproduction at moderate concentrations of the drug (10 to 15 μg ml^?1. (3) Resistance to 100 μg ml^?1 metronidazole, unstable in absence of selective pressure of the drug. (4) Resistance to high concentrations of metronidazole, stable when the organisms were maintained under nonselective conditions. The trichomonads with fully developed resistance were able to grow in anaerobic culture at 100 μg ml^?1 metronidazole and could be maintained indefinitely under these conditions. The minimal lethal concentrations for metronidazole obtained with these strains in an anaerobic in vitro assay were, at 48 h, 500 to 1000 μg ml^?1. This is 100 to 400 times higher than those obtained with the parent clones. The fully developed resistance was stable in organisms maintained in the absence of the drug over 2 years. The substrains with unstable resistance regained the susceptibility to high concentrations of metronidazole after 80 to 100 transfers in drug-free media. These strains, however, retained their resistance to moderate doses of metronidazole and full resistance could be restored by subculture in the presence of 10 μ ml^?1 metronidazole.     

Tritrichomonas foetus: Localization of filipin-sterol complexes in cell membranes

The polyene antibiotic, filipin, was used as a probe for the detection of sterols in the freeze-fractured plasma membrane and the flagellar membranes of the pathogenic protozoa, Tritrichomonas foetus. A homogeneous distribution of filipin-sterol complexes was seen throughout the plasma membrane, and the membrane of the three anterior and the one recurrent flagella. No or very few filipin-sterol complexes were observed in some specialized regions such as the base of the flagella (necklace), the portion of the recurrent flagellum, and that part of the cell body to which the flagellum was attached. The density of filipin-sterol complexes varied from one cell to the other. In some cells, about 205 complexes/μm² were seen. A larger number of filipin-sterol complexes were observed on both faces of the membrane of cytoplasmic structures, probably corresponding to vacuoles. No complexes were seen in the nuclear membrane and in the membrane of the endoplasmic reticulum. Very few or no complexes were observed in the membrane of the hydrogenosomes. Treatment of living cells with filipin induced aggregation of filipin-sterol complexes at some points of the plasma membrane.     

Trypanosoma cruzi: Isolation and characterization of a proteinase

Lysates of Trypanosoma cruzi epimastigotes were able to hydrolyze casein (K_m = 2.5 mg/ml) as well as bovine and human hemoglobins (K_m = 12.2 mg/ml); there was optimum activity was around pH 7.0. The proteinase activity detected with these substrates was enhanced by sodium diaminotetraacetate (EDTA) and reducing agents (SO^2?₃, mercaptoethanol, cysteine) and was inhibited by sulfhydryl reagents, thus suggesting an SH-dependent enzyme. Purification (60×) of the proteinase was carried out as follows: (1) precipitation at ?20 C, pH 4.5, with 80% acetone, (2) gel filtration on Sephadex G-200, (3) affinity chromatography on Sepharose 4B covalently linked to p-aminophenyl mercuric acetate. Only a single component (with an estimated molecular weight of 60,000) was detected in purified preparations by polyacrylamide gel electrophoresis. However, in addition to the major component identified as a proteinase, crossed immunoelectrophoresis experiments indicated the presence of at least three other antigens that apparently were devoid of proteinase activity. Optimum pH activity of the purified preparations was around pH 6.0 for casein and pH 3.0 for hemoglobins, but these activities probably are due to the one enzyme since they were altered identically by the same agents.     

Phocanema decipiens: Pathology in seals

Infective larvae of the anisakine nematode, Phocanema decipiens from cod (Gadus morhua), were fed to harbor seals (Phoca vitulina) and gray seals (Halichoerus grypus). At termination of the experimental infections, 6 of 8 harbor seals were infected with 34(6–64) P. decipiens, while the remaining 2 seals had relatively heavy infections of 250 and 547 nematodes, respectively; 10 of 11 gray seals were infected with 128 (68–229) P. decipiens, but only 2 nematodes were recovered from the 11th seal. Larvae and adults of P. decipiens occurred throughout the gastrointestinal tract, but mainly in the fundic portion of the stomach; the anterior extremities of the nematodes were embedded in mucosa and submucosa. Clusters of adult P. decipiens were associated with ulcerous gastric lesions in harbor seals and raised inflammatory areas in stomachs of gray seals. Singly occurring larvae from challenge transmissions were associated with raised inflammatory areas in the stomachs of both host species. Histological examinations revealed that the lesions and inflammatory areas were eosinophilic granulomata. Anisakis sp. larvae from the viscera of cod were also fed to 1 of the gray seals. Eighty of these larvae were subsequently found in association with a general inflammation in the cardiac area of the stomach in this seal. Natural anisakine infection were surveyed in 16 harbor and 53 gray seals from the Nova Scotia mainland. The natural incidence of P. decipiens was 62(5–177) in harbor seals and 577(11–1694) in gray seals. Clusters of adult P. decipiens were found in association with gastric lesions in 2 juvenile harbor seals; however, in gray seals, the nematodes neither occurred in clusters nor in association with gastric lesions.     

Nippostrongylus brasiliensis: Effect of cyclophosphamide treatment on worm expulsion in rats

The precise immunological mechanisms associated with expulsion of the gastrointestinal nematode Nippostrongylus brasiliensis remain controversial. In order to investigate the effects of drug-induced immunosuppression on parasite burdens and expulsion, various regimens of cyclophosphamide were administered to parasitized Wistar rats. It was observed that both the number of worms established from an infective dose of 3000 larvae and the time of expulsion were markedly increased with higher doses of cyclophosphamide. Thus, at the highest sublethal level of treatment (100 mg/kg), 82% of the infective dose was recovered at Day 9 postinfection compared with 51% in nontreated controls. Furthermore, in such treated rats expulsion was delayed in 6 days beyond that of nontreated animals. As cyclophosphamide, at the levels used in the present study, is known to primarily effect B-cell function, the results support the view that antibody-mediated responses play an essential role in worm expulsion.     

Litomosoides carinii: Effect of splenectomy on the ability of naive Mastomys natalensis to accept transplanted adults

Fifty-eight male rats Mastomys natalensis, were used in each of two experiments to study the effect of splenectomy on the rejection of transplnted nematodes, Litomosoides carinii. The rats were divided into five groups: splenectomized recipients (SR), normal recipients (NR), splenectomized (S), normal (N) and donor (D). Each group had 12 animals except for the D group which had 10. After patency of the D group (tank infected with Litomosoides carinii), groups SR and S were splenectomized. Fourteen days later, (three to five L. Carinii derived from the D group) were surgically transplanted into the peritoneal cavity of the SR and NR groups. In the first experiment, weekly sacrifices were made starting at Day 3. In the second experiment, all groups were sacrificed at Day 32. Worms transplanted into the SR group were accepted while those transplanted into the NR group were rejected. Sequentially examined antibody titers after Day 3 fell into two groups, those that were recipients of transplants (SR and NR) and those that were not (NR and SR). After becoming positive on Day 3, the microfilaremia of the SR group rose by Day 31 while that of the NR group fell to near 0. It was concluded that the spleen is necessary for the rejection of transplantedL. carinii by naive M. natalensis.     

Heligmosomoides polygyrus: Simple recovery of post-infective larvae from mouse intestines

Mice infected orally with third-stage larvae of Heligmosomoides polygyrus were killed at various times after infection. Their small intestines were removed, tied at each end and incubated at 37 C in dilute culture medium. When intestines were taken from mice infected for a period of between 1 and 7 days, a number of developing larvae comprising up to 20% of the infective dose emerged within 60 min through the intestinal wall into the medium. The recovery of emergent larvae was highest using intestines from mice infected 36 to 120 hr previously. The proportion of parasites emerging from the intestines of 48-hr-infected mice was similar for doses of 100 to 2400 larvae. Significantly fewer larvae emerged from the intestines of mice resistant to reinfection and challenged with third-stage larvae 36–72 hr before necropsy.     

Plasmodium berghei: The in Vitro immune response

Following primary in vitro Stimulation by Plasmodium berghei, IgM titers by the indirect fluorescent antibody test (IFAT) were negative in in vitro reconstituted syngeneic mouse spleen cultures containing T cells and macrophages, or B cells and macrophages, or macrophages alone, but IgM titers of 1:20 were obtained from cultures containing B cells, T cells, and macrophages. IFAT IgG titers were negative for cultures with T cells and macrophages together, or macrophages alone, but rose to 1:40 with cultures containing B cells and macrophages and 1,80 with cultures of B cells, T cells, and macrophages together. After a second in vitro challenge, IgM and IgG titers were similar to the stimulated cultures from immunized mice; the IgM titer reached only 1:20 but the IgG titer rose to 1:160. Total immunoglobulin was higher in the cultures from immunized mice than from in vitro primed cultures.