Evaluation of different agroforestry tree species for their suitability in the phytoremediation of seleniferous soils

Seven agroforestry tree species were grown in a clay loam soil treated with different levels of selenate‐Se, viz. 0, 1.25, 2.5 and 5.0 mg/kg supplied through sodium selenate. After 1 year of growth, a progressive decrease in dry matter of leaves, stem and roots was observed with increasing levels of applied Se. However, a significant decrease in dry matter yield was observed only at or above 2.5 mg Se per kg soil and shisham (Dalbergia sissoo) proved to be highly sensitive to the presence of selenate‐Se in the soil. On average, the largest above‐ground and below‐ground biomass was accumulated by arjun (Terminalia arjuna) and the lowest by the acacia tree (Acacia tortillas). The selenium content of leaves, stem and roots of all the tree species increased significantly with increasing levels of applied Se, although a large variation within species was observed. In the stem portion of different trees, the highest concentration of Se was found in dek (Melia azedarach) (5.1 mg/kg) and the lowest in mulberry (Morus alba) (2.6 mg/kg). The efficiency of selenium removal (including leaves, stem and roots) was the highest in arjun followed by eucalyptus (Eucalyptus hybrid) – Clone 10, mulberry, jambolin (Syzygium cumini), dek, shisham and acacia. Effective removal of Se takes place through the stem portion of different trees where it constitutes 30–50% of total Se. Large variation in Se uptake by different tree species suggests that trees vary in their potential for phytoremediation of seleniferous soils. In one growing season, shisham aged 24 years, poplar (Populus deltoides)– Clone G 48 (10 years old) and eucalyptus – Clone 10 (10 years old) could remove 2385, 1845 and 1407 g Se per hectare respectively. Corresponding reductions in Se capital of the soil varied between 24 and 37, 19 and 29 and 14 and 32%, respectively, in the surface layer (0–15 cm) alone or 7–11, 6–9 and 4–7% for the whole soil profile (0–120 cm). Removal further increased to 4207 g Se per hectare under an agroforestry farming system of poplar–mentha/wheat with Se being reduced from 43 to 65% for the surface layer and from 13 to 20% for the whole profile.     

Effects of non-chemical soil fumigant treatments on root colonisation with arbuscular mycorrhizal fungi and strawberry fruit production

The effects of biofumigation and soil heating on arbuscular mycorrhizal fungi (AMF) colonisation, strawberry growth and strawberry yield in pot experiments compared with untreated soil and chemical fumigation with dazomet were tested. Three different Brassicaceae species (Brassica juncea, Eruca sativa, Sinapis alba) were used as biofumigant plant green manure and soil heating was applied to simulate soil solarisation. Half of the plants were inoculated with indigenous arbuscular mycorrhizal fungi inoculum. With one exception (E. sativa) among the uninoculated plants, the treatments significantly decreased the mycorrhizal colonisation parameters compared with the untreated control. Dazomet displayed the greatest inhibitory effects on AMF establishment. In addition, the intensity and number of bands corresponding to Glomus spp. obtained with temporal temperature-gradient gel electrophoresis were lower for strawberry plants from biofumigant treatments than from the control. For the inoculated plants, there were almost no significant differences among the mycorrhizal colonisation parameters. The mass of leaves for the uninoculated and inoculated plants was higher for almost all non-chemical soil fumigant treatments compared with the control, except for heating of the uninoculated treatments. The number of strawberry fruits for the uninoculated biofumigant treatments was the highest, being higher than the values observed for the heating treatments, the chemical disinfection treatments and the control. There were no significant differences among the inoculated treatments. Biofumigation with Brassicaceae species resulted in higher soil organic matter and mineral nutrients and had a relatively small effect on AMF colonisation (F% = 59.0, 80.3, 47.3 for Bj, Es and Sa, respectively) compared with uninoculated controls (F% = 84.3). Despite the reduced AMF colonisation, biofumigation resulted in a higher fruit number and mass of leaves. Therefore, it represents a non-chemical soil fumigation method that should be applied in sustainable strawberry production.     

Development of molecular markers to assess the level of introgression of Populus tremula into P. alba natural populations

Morphological traits traditionally adopted to discriminate between Populus alba L. and P. tremula L. have frequently led to misclassifi‐cation of their spontaneous hybrid P. × canescens Sm. Moreover, they may not be of any help in cases of spontaneous backcross phenomena. These limitations can be overcome by molecular markers, which are not environmentally influenced nor subjectively assessed. In this study, the effectiveness of amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers in species and hybrid discrimination was evaluated by analysing a set of reference samples of P. alba, P. tremula and P. × canescens. Species‐specific and species‐indicative AFLPs, as well as diagnostic SSR alleles, were recorded in both P. alba and P. tremula reference samples. The results allowed a clear distinction between the two poplar species and their hybrid. Using these diagnostic markers, a natural population of P. alba trees sampled along the Ticino river basin in northern Italy was analysed, and P. × canescens individuals, intermingled with P. alba trees, were detected.     

转透明颤菌血红蛋白基因杨树的再生及其生长量初步测定

采用农杆菌介导法,将透明颤菌血红蛋白Vitreoscilla hemoglobin(VHb)基因vgb导入银腺杨(Populus alba ×P.glandulosa).以经无菌培养的叶片为外植体,通过450个叶盘与根癌农杆菌(Agrobacterium tumefaciens)LBA4404共培养,将植物双元表达载体pPZV中vgb基因导入银腺杨基因组,经卡那霉素筛选后,共获得60株卡那霉素抗性植株.经PCR特异性扩增和Southern点杂交分析,证明其中52株再生植株基因组DNA中整合了vgb基因.RT-PCR分析结果表明,vgb基因在其中的44个株系中获得表达.在2年温室生长条件下,转基因无性系的外部形态与对照相比无显著差异.但在株高和地径上,有3株转基因无性系较对照植株有明显增加.     

落葵红色素作为酸碱指示剂的研究

以落葵果为原料提取红色素,测定落葵红水溶液的变色pH值为8.5～10.4,与酚酞相同,且变色灵敏,可以作为酸碱滴定指示剂并成为酚酞指示剂的代用品。用落葵红与酚酞一起作为对食品酸味剂和膨松剂分析指示剂的实验结果表明,落葵红指示酸碱滴定终点变色明显,与酚酞相当。落葵红指示剂宜现用现配,而制成的浸膏在无水乙醇密封下避光避热可长期保存。落葵红用作指示剂的安全性高,具有实用价值。     

河北杨和新疆杨离体叶片诱导不定芽研究

对河北杨和新疆杨离体叶片诱导不定芽的方法进行了研究。结果表明,适合河北杨和新疆杨离体叶片不定芽诱导的最适培养基为:1/2M S+0.25 m g/L 6-BA+0.01 m g/L TDZ+0.25 m g/L IAA;在此培养基中,叶片分化率河北杨可达70%,新疆杨可达52%;平均分化芽数河北杨为6.84个,新疆杨为5.32个。6-BA单独使用对叶片不定芽诱导没有效果,NAA作用不明显。叶片刻伤方式对两种杨树叶片诱导不定芽影响显著,沿中脉横切效果优于去除叶边缘,诱导的不定芽数前者比后者约多3个。光照显著影响叶片不定芽产生的时间,不定芽在3~5 d暗培养后转至16 h/d光照下培养比全程16 h/d光培养发生早15 d左右;光照对分化率和平均分化芽数无显著影响。     

桑树不同方位叶片中葫芦巴碱含量测定

为解决利用分光光度法和极谱法测定植物中葫芦巴碱含量时时间较长,以及样品的制备和提取比较繁琐,操作复杂等问题,建立了高效液相色谱法测定桑树不同方位叶片中葫芦巴碱的含量方法。以氨基键和柱为固定相,甲醇和水为流动相,柱温为室温,可快速准确测定桑树叶片中葫芦巴碱含量。本方法样品预处理简单、分析灵敏度高、精密度好、速度快,尤其适应批量样品的分析。结果表明,桑树东南叶片中葫芦巴碱含量较高,西北叶片中葫芦巴碱含量较少,这可能与光照强度有关。     

银中杨茎段离体培养再生植株研究

以银中杨茎段为外植体,建立组织培养系统.试验结果表明,MS为最适基本培养基;各激素对诱导不定芽的作用大小为TDZ＞6-BA＞KT,诱导不定芽的最佳培养基为MS+0.1 mg·L-1TDZ+蔗糖20 g·L-1+琼脂0.7%;诱导不定芽再生的最佳光质为白光或黄光;生根培养基以1/2MS+0.5 mg·L-1IBA+蔗糖2...     

三个桑树肌动蛋白基因的克隆与组织表达分析

肌动蛋白基因在植物各种生理活动中具有极其重要的作用,通过同源克隆与反向PCR的方法,克隆了3个肌动蛋白基因的核心片段,其中一个为已报道的MaACT1,另两个分别被命名为MaACT2和MaACT3,进而PCR扩增获得MaACT1和MaACT2肌动蛋白全长CDS,其中MaACT2基因全长1 704 bp,由4个外显子和3个内含子组成,CDS为1 134 bp,编码377个氨基酸残基。采用RT-PCR的方法分析了3个基因在叶、茎、果、根等组织的表达情况以及在茎、叶和托叶的生长过程中的表达变化。MaACT1在茎中表达量较弱但有随着茎的生长逐渐增强的趋势,在幼叶中有较高的表达,MaACT2与MaACT3在根、茎、叶等组织中都有较高表达,MaACT3在叶、托叶和茎的各个发育时期表达都很稳定,可以作为桑树基因表达研究的内参基因。