两种浮萍植物的叶绿体超微结构对模拟酸雨的敏感性

用蒸馏水和不同pH值(5.5,4.5,3.5和2.5)的模拟酸雨培养浮萍(Lemn aminorL.)和紫萍(Spirodela polyrrihiza(L.)Schleid)48h后,测定叶状体膜系统渗漏率,观察叶绿体超微结构的变化。结果表明,随酸雨pH的下降,两种浮萍叶状体的膜系统渗率增大,叶绿体超微结构受损。浮萍和紫萍对酸雨的敏感性和伤害时细胞与叶绿体形态变化显示一定的种间差别。浮萍的结构性损伤始于pH4.5,原生质体收缩,出现质壁分离,基粒结构混乱;pH3.5时叶绿体肿胀呈球状,片层结构破坏并出现许多空泡。紫萍在pH3.5时膜系统的外渗率仍较低,基粒和基质类囊体结构无明显改变;pH2.5时叶绿体结构才出现严重伤害,但未见明显的肿胀与质壁分离现象。因此认为在两种浮萍的共生水体中,紫萍对酸雨污染有较强的生存竞争能力,而浮萍则可用作pH<4.5水体的灵敏指示植物。     

紫萍营养繁殖过程的解剖观察

以采自海南陵水的紫萍(Spirodela polyrhiza DW2501-4)为材料,观察紫萍的营养繁殖过程。紫萍DW2501-4培养在0.5倍Hoagland′s液体培养基中,先不断长出新的叶状体,随着营养物质的减少,长出休眠体进入休眠状态。休眠体经过4℃处理7 d后,可在含1%蔗糖的Hoagland′s固体培养基中重新萌发。为了进一步观察叶状体、类休眠体和休眠体的组织结构差异,以及休眠体萌发过程中组织结构的变化,制作了石蜡切片。观察结果显示:紫萍叶状体有数条叶脉,细胞中含少量的淀粉粒,表皮层以下有分层的气室,通常分上下两层,上层气室比下层气室小。类休眠体也有数条叶脉和少量气室,细胞中有较多淀粉粒。休眠体的细胞差异不明显,几乎没有气室,细胞内有大量的淀粉粒,部分细胞含有单宁;随着休眠体萌发,细胞中淀粉粒不断变小,同时分生组织分化出根和新的叶状体。     

细胞分裂素在维持紫萍叶状体生命中的重要作用

将无根和囊的紫萍(Spirodela polyrrhiza)P143品系半叶状体分别放在含有和不含6-苄基嘌呤的培养液上于长日照条件下培养。在不含6-BA的培养液上培养4d以后,半叶状体开始失绿,光合能力降低;第12天时,半叶状体已接近死亡。但是,培养在含有6-BA的培养液上的半叶状体干重、叶绿素和可溶性蛋白质含量、希尔反应活力、核酮糖-1,5-二磷酸羧化酶/加氧酶的小亚基和其编码基因rbcS mRNA水平显著增加。这些6-BA处理过的半叶状体平均寿命可达80d,几乎等同于完整植物体上叶状体的寿命,因此提出细胞分裂素对于紫萍叶状体生物学功能的维持是必需的。     

纳米银对紫萍休眠芽萌发、存活和生长的影响

纳米银(AgNPs)是一种潜在的新型环境污染物,本研究以紫萍(Spirodela polyrhiza (L.) Schleid)休眠芽为实验材料,研究AgNPs对休眠芽萌发率、存活率及叶状体数目、面积和叶绿素含量等指标的影响,并对各项指标的半数效应浓度(EC₅₀)进行比较。结果显示,AgNPs可以抑制紫萍休眠芽的萌发,高浓度(10 mg/L)时可造成休眠芽死亡率显著增加。紫萍休眠芽萌发后叶状体数目和叶状体面积、色素含量均随浓度增加逐渐降低,表现出剂量效应,且叶绿素a对AgNPs最敏感。研究结果表明AgNPs对水生植物紫萍无性繁殖体的萌发和生长都具有抑制作用,具有一定的生态风险。     

紫萍叶状体衰老过程中的内肽酶谱变化和外源L-丝氨酸对内肽酶谱的影响

采用双向凝胶电泳技术分析紫萍叶状体衰老期间内肽酶同工酶的变化以及外源L-丝氨酸对内肽酶影响的结果表明,在衰老的紫萍叶状体中检测到9种内肽酶同工酶,丝氨酸内肽酶EP3可能在叶状体衰老的早期起作用,而半胱氨酸内肽酶EP9在第6天出现,是衰老后期活性最强的内肽酶。培养液中加入外源的L-丝氨酸后,叶状体蛋白质含量下降加剧,与衰老相关的内肽酶EP3、EP4和EP9的活性明显增强或提前出现。     

长江中游浮萍种质资源鉴定及遗传多样性分析

浮萍为浮萍科水生植物,具有生长繁殖快、氮磷吸收能力强及高含蛋白质和淀粉的特性,在饲料和能源方面具有较大的开发潜力。目前尚未有系统研究我国长江中游地区浮萍种质资源的报道。本研究从我国长江中游地区3个省42个具有代表性的自然水体中共收集到浮萍种质112份。经形态学并结合分子生物学鉴定,它们分属4个属6个种,其中青萍属(Lemna L.)含3个种54份种质;多根紫萍属(Spirodela Schleid.)、少根紫萍属(Landoltia LesCrawford)、芜萍属(Wolffia Horkel ex Schleid.)均只有1个种,种质数量分别为33份、19份和6份。遗传多样性分析表明Atp F-Atp H间隔序列和rp S16内含子序列的单倍型数量分别为11和18,单倍型多样性分别为0.748和0.892,群体突变率分别为0.03013和0.03507,平均每Kb核苷酸差异数为31.386和61.263,核苷酸多样性分别为0.04713和0.06623。本研究为我国长江中游浮萍的资源化利用奠定了基础。     

不同营养状态下金鱼藻的生理响应

通过静态模拟实验,比较研究了不同营养水平(中营养、富营养、重富营养和Hoagland植物培养液)下培养的金鱼藻(Ceratophyllum demersum)的蛋白质、叶绿素含量,过氧化物酶(POD)及超氧化物酶(SOD)活性变化.研究发现,总N、总P的变化会影响金鱼藻的生物合成,当水环境总氮浓度低于1 mg·L^-1,总磷浓度低于0.1 mg·L^-1时,金鱼藻茎叶的叶绿素合成较低,其茎蛋白含量迅速下降.金鱼藻在富营养水平(1 mg TN·L^-1,0.1 mg TP·L^-1)下抗氧化防御酶活跃,POD、SOD活性增高.研究表明,金鱼藻较适应于富营养水环境,水体营养盐继续增加对金鱼藻有胁迫作用,过高营养盐浓度(重富营养和Hoagland 培养液)影响金鱼藻的抗逆能力.     

日本青萍6746的种质保存

日本青萍6746(Lemna paucicostata 6746)是用作光周期诱导成花实验的理想材料,国内各大学已广泛采用,周有院等对这方面的实验设计和研究结果曾做过详细报道。目前对日本青萍6746的种质保存普遍采用叶状体继代培养方法,即将3～4片叶状体接种在液体或固体培养基上,让其出芽繁殖,但需经常转接,以达到种质保存的目的。如果半年,     

紫萍P143品系植物rbcS基因的cDNA克隆与表达

紫萍P14 3品系离体半叶状体在黑暗下培养 4d以后 ,rbcS基因的表达比完整植株显著降低 ;半叶状体在黑暗下培养 10d ,后再转移到长日照条件下培养 ,rbcS基因的表达能恢复到较高水平     

营养液强度对AM真菌生长发育的影响*

研究了在贫营养基质中不同强度Hoagland营养液对丛枝菌根(Arbuscular mycorrhizae, M)真菌Glomus versiforme生长发育的影响。结果表明:本试验条件下,菌根侵染率、菌丝量、孢子数间呈显著正相关。在施加5%~50%强度Hoagland营养液时, 菌根真菌的生长与宿主植物高粱根中磷浓度、可溶性糖浓度密切相关,而与氮浓度无显著相关。由此认为:在盆栽生产菌根菌剂时,基质中存在一个临界磷浓度,在这个临界浓度之下,菌根真菌的生长发育随磷浓度的提高而增长,超过该临界浓度则会随磷浓度的提高而下降。施用20%、50%强度Hoagland营养液对菌根真菌生长最为有利,其菌根侵染率、菌丝量、孢子数均高于其它处理,因此认为:宿主植物—菌根真菌之间共生关系的基础是营养条件,基质中养分的高低会影响互惠共生关系的建立和发展。在高质量菌剂生产中, 菌根共生体双方的生长发育完全可以由人工控制。施加营养液是一种有效的调控手段,有可能使共生平衡向有利于菌根真菌生长发育的方向倾斜,使真菌得到最大程度的生长。     

Ammonium and nitrate uptake by the floating plant Landoltia punctata

BACKGROUND AND AIMS: Plants from the family Lemnaceae are widely used in ecological engineering projects to purify wastewater and eutrophic water bodies. However, the biology of nutrient uptake mechanisms in plants of this family is still poorly understood. There is controversy over whether Lemnaceae roots are involved in nutrient uptake. No information is available on nitrogen (N) preferences and capacity of Landoltia punctata (dotted duckweed), one of the best prospective species in Lemnaceae for phytomelioration and biomass production. The aim of this study was to assess L. punctata plants for their ability to take up NH4+ and NO3- by both roots and fronds. METHODS: NO3- and NH4+ fluxes were estimated by a non-invasive ion-selective microelectrode technique. This technique allows direct measurements of ion fluxes across the root or frond surface of an intact plant. KEY RESULTS: Landoltia punctata plants took up NH4+ and NO3- by both fronds and roots. Spatial distribution of NH4+ and NO3- fluxes demonstrated that, although ion fluxes at the most distal parts of the root were uneven, the mature part of the root was involved in N uptake. Despite the absolute flux values for NH4+ and NO3- being lower in roots than at the frond surface, the overall capacity of roots to take up ions was similar to that of fronds because the surface area of roots was larger. L. punctata plants preferred to take up NH4+ over NO3- when both N sources were available. CONCLUSIONS: Landoltia punctata plants take up nitrogen by both roots and fronds. When both sources of N are available, plants prefer to take up NH4+, but will take up NO3- when it is the only N source.     

Differential sensitivity of four Lemnaceae species to zinc sulphate

Lemnaceae are currently the only freshwater plants required for regulatory toxicity testing of pesticides and other chemicals. Toxicological protocols allow for the use of different Lemnaceae species in tests. However, few studies have compared the relative sensitivity of individual duckweed species. Zinc is an essential plant nutrient but is also a common pollutant in aquatic environments and elevated levels are phytotoxic. This study shows that four species of Lemnaceae differ in their relative sensitivities to zinc sulphate, a commonly used reference chemical. Comparative zinc sensitivity, in order, from most tolerant to most sensitive was: Landoltia punctata > Lemna minor > Wolffia brasiliensis > Lemna gibba. Zinc sensitivity was also endpoint dependant. EC₅₀ values typically increased in order of: specific biomass growth rate < specific frond number growth rate < chlorophyll absorbance. However, specific frond number growth rate was the most sensitive endpoint for L. punctata. Unlike the other species, L. punctata displayed no significant colony disintegration. Lemna species and L. punctata appear to be employing distinct response strategies when exposed to zinc. L. gibba and L. minor produce and release young, single fronds which are severely affected by zinc. In contrast, L. punctata produces fewer fronds, which are not released and form large colonies of high biomass that are relatively zinc tolerant.     

SELF‐THINNING AND SIZE INEQUALITY DYNAMICS IN A CLONAL SEAWEED (SARGASSUM LAPAZEANUM,PHAEOPHYCEAE)1

Fronds of clonal seaweeds with extensive holdfasts relative to frond size are known not to self‐thin during growth, even in crowded stands. We tested whether frond self‐thinning would occur for such a seaweed since these traits are more similar to those of unitary seaweeds, which do self‐thin in crowded conditions. We used Sargassum lapazeanum Setch. et N. L. Gardner (Fucales, Phaeophyceae) from the Pacific coast of Mexico, for which we first confirmed its clonal nature by performing a regeneration experiment in culture tanks. During the growth season (winter to late spring), S. lapazeanum stand biomass increased, while frond density and size inequality (Gini coefficient for frond biomass) decreased. These results indicate that self‐thinning occurred at the frond level. We propose a conceptual model for frond dynamics for clonal seaweeds in general. In stands of clonal species with small fronds and relatively extensive holdfasts (particularly when holdfasts are perennial), frond dynamics would be determined mostly by intraclonal regulation, which seems to prevent excessive crowding from occurring. Such species display a positive biomass–density relationship during the growth season. On the contrary, in stands of clonal species with large fronds relative to holdfast size, frond dynamics would be determined mostly by interactions among genets. For such species, self‐thinning may be detected at the frond level in crowded stands, resulting in a negative biomass–density relationship during growth.     

Rapid and highly efficient callus induction and plant regeneration in the starch-rich duckweed strains of <Emphasis Type="Italic">Landoltia punctata</Emphasis>

The starch-rich duckweed Landoltia punctata is a valuable aquatic plant in wastewater purification, bioenergy production, and many other applications. A highly efficient callus induction and plant regeneration protocol is desirable so that biotechnology can be used to develop new varieties with added value and adaptation. We studied both known and unknown factors that influence callus induction in L. punctata and obtained almost 100 % induction rate in 30 days. The optimum medium for callus induction was MS basal medium supplemented with 1 % sorbitol, 15 mg/L 2,4-D, and 2 mg/L 6-BA. Green fragile callus was induced from the meristematic region in the budding pouches. The optimum photoperiod for callus induction was 16-h day, and the optimum explant orientation was dorsal side down on the medium. The optimum medium for callus subculture was WPM basal medium supplemented with 2 % sorbitol, 4 mg/L 2,4-D, and 0.5 mg/L TDZ. Green callus could be maintained by subculture once every 4 weeks. However, when the subculture cycle was prolonged to 6 weeks or longer, yellow fragile embryogenic callus was obtained. The optimum plant regeneration medium was MS medium supplemented with 0.5 % sucrose, 1 % sorbitol, and 1.0 mg/L 6-BA with frond regeneration rates of approximately 90 %. The regenerated fronds rooted in Hoagland’s liquid medium in 1 week. The callus induction and frond regeneration protocol was tested for its efficiency in geographically distinct strains 5502, 8721, and 9264. Thus, we obtained a rapid and efficient protocol for callus induction and frond regeneration of L. punctata, which takes only 9 weeks.     

Regeneration and reproduction of Mazzaella cornucopiae (Rhodophyta, Gigartinaceae) after frond harvesting

The effects that different intensities of frond harvesting have on frond regeneration and subsequent production of reproductive structures were investigated for the red intertidal alga Mazzaella cornucopiae (Postels & Ruprecht) Hommersand from British Columbia, Canada. Harvesting was done by pruning fronds in the late spring (when stand biomass is highest) of 1993 at two intensities: total and partial collection of fronds, in this second case leaving all frond biomass less than 1 cm high in place. Holdfasts were not damaged. Total percent cover of thalli, frond density, mean frond length, and stand biomass for these experimental quadrats were statistically similar to values for control quadrats in the spring of 1994. These results suggest that one total harvest of fronds per year, done in late spring without damaging holdfasts, may give the highest sustainable yield of biomass. The effects of harvesting intensity on reproduction were variable and difficult to explain. Neither the appearance nor the abundance of cystocarpic fronds were affected by frond pruning, compared with control areas, but pruning did affect the appearance and the abundance of tetrasporic fronds. Partial pruning resulted in a longer presence of tetrasporic fronds, whereas total pruning was associated with their complete absence. Results are compared with those for the few other species of the Gigartinaceae for which experimental harvesting has been done. This revised version was published online in August 2006 with corrections to the Cover Date.     

Interaction of arsenic and phosphate on their uptake and accumulation in Chinese brake fern

Interactive effects of arsenate (As (V)) and phosphate (Pi) were investigated under hydroponic culture. Arsenic concentrations in fronds and roots of Chinese brake fern (Pteris vittata L.) significantly (p < 0.05) increased with increasing As (V), but decreased (p < 0.05) with increasing Pi in nutrient solution. Phosphate uptake was significantly (p < 0.05) inhibited by 1000 micromol L(-1) As (V). Under 100 micromol L(-1) As (V), frond phosphorus (P) increased at 100 and 1000 micromol L(-1) Pi, and root P increased at 250 micromol L(-1) Pi exposures. Arsenic and P concentrations in fronds and roots of Chinese brake fern were negatively correlated (p < 0.05). Arsenate treatments enhanced As and P transport to fronds, while increasing Pi inhibited their transportation, with highest frond P and As (%) obtained under 100 micromol L(-1) treatment. pH values in nutrient solution increased with increasing exposure time, but decreased with increasing Pi levels. Dissolved organic carbon (DOC) contents (dry weight) in nutrient solution decreased with increasing Pi levels, both for treatments with and without As (V). Arsenate at 1000 micromol L(-1) significantly (p < 0.05) increased DOC contents, especially for treatment without Pi. Six organic acids were detected in root exudates of Chinese brake fern, with oxalic and malic acids being most dominant.     

Growing duckweed in swine wastewater for nutrient recovery and biomass production

Spirodela oligorrhiza, a promising duckweed identified in previous studies, was examined under different cropping conditions for nutrient recovery from swine wastewater and biomass production. To prevent algae bloom during the start-up of a duckweed system, inoculating 60% of the water surface with duckweed fronds was required. In the growing season, the duckweed system was capable of removing 83.7% and 89.4% of total nitrogen (TN) and total phosphorus (TP) respectively from 6% swine lagoon water in eight weeks at a harvest frequency of twice a week. The total biomass harvested was 5.30 times that of the starting amount. In winter, nutrients could still be substantially removed in spite of the limited duckweed growth, which was probably attributed to the improved protein accumulation of duckweed plants and the nutrient uptake by the attached biofilm (algae and bacteria) on duckweed and walls of the system.     

A novel mechanism of abscission in fronds of Lemna minor L. and the effect of silver ions

Lemna minor L. (duckweed) forms colonies through vegetative propagation because mother fronds remain connected for some time with their daughter fronds by stipes. The colony size is controlled by abscission of stipes at a specific preformed abscission zone. Application of silver ions (Ag(+) ) enhances the rate of frond abscission, thus resulting in smaller colonies. The mechanism behind this process has not yet been identified. Silver caused an abscission response that saturated after 7 h of treatment. The half-maximal effective concentration was 0.72 μm Ag(+) for the standard clone, L. minor St. Other clones of the same species show sensitivities that differ by one order of magnitude. Transmission electron microscopy revealed: (i) large numbers of vesicles close to the plasmalemma in cells adjacent to the abscission zone, which proves a vesicular type secretory activity; and (ii) a moderately electron-dense secretion accumulated in the enlarging intercellular spaces, and seemed to flow from the adjacent cells towards the abscission zone. We assume that increasing pressure causes this material to push apart the cells, thereby causing the break in the abscission zone of the stipe. This is a novel mechanism of abscission that has not previously been described. The same mechanism occurs in stipes of both control and Ag(+) -treated samples. Silver ions only accelerate the process leading to abscission of stipes, without affecting the mechanism involved.     

pH及无机氮化合物对细脉浮萍生长的影响

研究以细脉浮萍 ( Lemna aequinoctialis)为研究对象 ,在完全培养液的基础上模拟生活污水的氨氮、硝酸氮浓度和 p H范围 ,研究了培养条件对其生长的影响。结果表明 ,这个浮萍种对 p H耐受的低限在 4～ 5之间 ,最适 p H范围为 5～ 6。非离子态的氨会对其生长产生抑制作用 ,在氨氮为唯一氮源的条件下 ,非离子态氨的浓度大于 0 .1 mg/L时会对此种的生长产生明显的抑制作用 ,接近 2 mg/L时 ,基本上致死。在研究的氨氮和硝酸氮浓度范围内 ( <40 mg N /L) ,铵根离子和硝酸根离子浓度增加可促进细脉浮萍的生长。氨氮中生长的植株过氧化物酶活性高于硝酸氮中的活性。