Optimum nitrogen fertilization of Calophyllum inophyllum seedlings under greenhouse conditions

A greenhouse pot experiment was conducted to study the effects of nitrogen fertilization on Calophy- llum inophyllum seedlings grown with 0, 50, 100, 150, 200, 300, 400 and 600 mg N per seedling according to exponential functions. Seedling height, root collar diameter, leaf area and total biomass increased with increasing fertilization from 0 to 200 mg N per seedling and decreased with further increase in fertilization from 300 to 600 mg N per seedling. The net photosynthetic rate, stomatal conductance, intercellular CO₂ concentration and transpiration rate of C. inophyllum seedlings showed a unimodal parabolic trend, with peak values of 7.29 mmol·m⁻²·s⁻¹, 0.071 mol·m⁻²·s⁻¹, 220 mmol·mol⁻¹ and 1.34 mmol·m⁻²·s⁻¹, respectively, when the rate of fertilization was 200 mg N per seedling. Photosynthetic gas exchange parameters were significantly different among nitrogen treatments. Based on the critical values of leaf N and P concentration and N/P ratio, the optimum amount of nitrogen of C. inophyllum seedlings was 200–400 mg per seedling for leaf N and P concentration, and 100–400 mg per seedling for N/P ratio. It was concluded that 200–400 mg N per seedling was the most suitable nitrogen range for C. inophyllum seedlings.     

Synthesis and anticancer activity of (+)-nopinone-based 2-amino-3-cyanopyridines

Twelve (+)-nopinone-based 2-amino-3-cyanopyridines 4a–l were synthesized from (–)-β-pinene. The structures of these compounds were characterized by FT-IR, ¹H NMR, and ESI-MS. All the compounds were tested for their anticancer activity against lung cancer cell line A549, gastric cancer cell line MKN45 and breast cancer cell line MCF7 by MTT method, respectively. The results showed that compounds 4f, 4j and 4k had promising anticancer activity against these cancer cell lines, in particular, compound 4f exhibited broad-spectrum and highly efficient anticancer activity against cell lines A549, MKN45 and MCF7 with IC₅₀ of 23.78, 67.61 and 53.87 µmol·L⁻¹, respectively. The preliminary analysis of the structure activity relationship implied that the Br or Cl substituted group of the benzene ring in these derivatives significantly contributed to the anticancer activity.     

A comparison of the biological activities of ethyl acetate fractions from the stems and leaves of Penthorum chinense Pursh

Penthorum chinense Pursh (PCP) is a popular traditional medicinal plant in China, widely used for the treatment of a variety of liver diseases. Although it has been long recognized that the main active elements of PCP are contained in ethyl acetate fraction (EAF), little is known so far in terms of the relative effectiveness of EAF derived from the stems versus leaves of this plant. In the current study, we prepared EAF by reflux extraction and sequential extraction from the stems (SEAF) and leaves (LEAF) of PCP and tested their hepatoprotective efficacies. The extract rates and flavonoid contents of LEAF were higher than those of SEAF. EAFs (>50 μg·mL⁻¹) prevented lipid accumulation in cells and protected against lipotoxicity injury when the concentration exceeded 25 μg·mL⁻¹. More than 95% free radicals released by 2,2-diphenyl-1-picrylhydrazyl (DPPH) were eliminated by 25 μg·mL⁻¹ SEAF and 50 μg·mL⁻¹ LEAF, respectively. Further, EAFs (25 μg·mL⁻¹) also showed protective antioxidant effects, with the activity of LEAF being significantly higher than that of SEAF. EAFs (10 mg·mL⁻¹) also showed similar unspecific bacteriostatic activity. In comparison with SEAF, LEAF contained more flavonoids and had a higher anti-oxidation capability and for these reasons we suggest it should be better for clinical use.     

2苹果品种叶片胚状体再生体系的建立

以苹果“富士”和“鲁加6号”为材料,建立叶片胚状体再生体系。结果表明:先经胚性细胞诱导培养基MS+BA1.0 mg·L^-1+NAA5.0 mg·L^-1+TDZ0.5 mg·L^-1诱导胚性细胞,再置于胚胎发育培养基MS+BA1.0 mg·L^-1+NAA0.1 mg·L^-1上进行暗培养,最佳时间为10 d,2品种叶片再生率分别为96.7%和100%。经胚状体不同时期形态学组织学观察,显示胚状体发育时期经历原胚、球形胚、心形胚、子叶形胚、鱼雷形胚、成熟胚等阶段。“鲁加六号”苹果最佳生根培养基为1/2MS+IAA0.1 mg·L^-1+IBA0.1 mg·L^-1,生根率达86.7%。     

富士和嘎啦苹果叶片面积估算模型构建

【目的】建立准确、无损的适宜于苹果不同品种和枝梢类型的叶面积估算模型。【方法】以富士及嘎啦不同长度的营养枝梢和果台枝梢叶片为试材,采用数字扫描仪获取叶片长度(LL)、宽度(LW)和叶面积(LA)等叶片形态参数,并采用决定系数(R²)、均方根误差(RMSE)、赤池信息准则(AIC)对建立的17个有常数项和无常数项叶面积模型精度进行筛选和适宜性评价。【结果】共获得5 207枚叶片形态参数,其中叶面积变异系数最大,达51.59%。叶片形态受品种及枝梢类型的显著影响,其中长枝梢叶片长、宽和叶面积显著大于同类型短枝梢叶片,而营养枝梢叶片长、宽和叶面积显著大于同长度果台枝梢,嘎啦叶片相比富士更为细长。以LL和LW复合变量为自变量的模型5:LA=a(LL×LW)、模型6:LA=a(LL+LW)²、模型9:LA=aLL²+bLW²、模型16:LA=a(LL×LW)^b、模型17:LA=(LL×LW)^b的精度可满足富士和嘎啦各类枝梢叶面积的估算,但需针对各品种和枝梢类型单独建...     

A novel aldo-keto reductase gene, IbAKR, from sweet potato confers higher tolerance to cadmium stress in tobacco

High concentrations of Cd can inhibit growth and reduce the activity of the photosynthetic apparatus in plants. In several plant species, aldo-keto reductases (AKRs) have been shown to enhance tolerance to various abiotic stresses by scavenging cytotoxic aldehydes; however, few AKRs have been reported to enhance Cd stress tolerance. In this study, the gene IbAKR was isolated from sweet potato. The relative expression levels of IbAKR increased significantly (approximately 3-fold) after exposure to 200 mmol·L⁻¹ CdCl₂ or 10 mmol·L⁻¹ H₂O₂. A subcellular localization assay showed that IbAKR is predominantly located in the nucleus and cytoplasm. IbAKR-overexpressing tobacco plants showed higher tolerance to Cd stress than wild-type (WT). Transgenic lines showed a significant ability to scavenge malondialdehyde (MDA) and methylglyoxal (MG). In addition, proline content and superoxide dismutase activity were significantly higher and H₂O₂ levels were significantly lower in the transgenic plants than in the WT. Quantitative real-time PCR analysis showed that the reactive oxygen species (ROS) scavenging genes encoding guaiacol peroxidase (GPX), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR) and peroxidase (POD) were significantly upregulated in transgenic plants compared to WT under Cd stress. These findings suggest that overexpressing IbAKR enhances tolerance to Cd stress via the scavenging of cytotoxic aldehydes and the activation of the ROS scavenging system.     

Phosphorus status,use and recycling in a Chinese peri-urban region with intensive animal husbandry and cropping systems Results from case study in a Sino-German applied research collaboration project

The Sino-German research collaboration project, “Recycling of organic residues from agricultural and municipal origin in China” (2008–2012), comprising different interdisciplinary research groups, and also German small and medium-sized enterprises, aimed at developing integrated strategies and solutions for the recycling of organic residues in China. In an intensive crop-livestock agricultural region in the Shunyi District of Beijing, five typical cropping systems were investigated. The research was conducted in the form of analyses of phosphorus (P) in soil, plants, animal feed, animal products, manures, mineral and organic fertilizers and the derivation of the corresponding nutrient balances and P flows. The mean annual P balance surplus was 492 kg·ha⁻¹·yr⁻¹ P for the vegetable production system, significantly higher (P<0.05) than that for orchards (130 kg·ha⁻¹·yr⁻¹P) and cereal crops (83 kg·ha⁻¹·yr⁻¹ P). Plant-available P (Olsen-P) concentrations of topsoils (0–20 cm) had good correlations with the amounts of P applied (from mineral and organic sources). Compared to results from the Second Chinese National Soil Survey of 1981, mean concentrations of available P in soils of 19 plots investigated in Shunyi District increased 10-fold (from 7.3 to 60 mg·kg⁻¹) from 1981 to 2009. On average, the critical limit for Olsen-P concentrations (>30 mg·kg⁻¹) that can lead to increased risk of P loss was exceeded in all five cropping systems. With feed additives, the “natural background value” (Chinese Environmental Quality Standard for Soils) of copper and zinc in topsoils was exceeded at several sites. Screening for several substances in the veterinary antibiotic classes of sulfonamides, tetracyclines, and fluoroquinolones revealed widespread topsoil contamination. Calculated livestock densities were 10.6 livestock units per ha arable land in 2007. Animal husbandry is increasingly conducted in large operations, making traditional ways of reuse difficult to apply. Comparing three management systems for treatment of organic residues from a pig farm via aerobic (composting) or anaerobic (biogas) treatment in a life cycle assessment, the resulting cropland demand for a sustainable land application of biogas effluent varied between 139 and 288 ha·yr⁻¹, well above the cropland area owned by the farm (10 ha). The mismatch problems in the above context between business-as-usual and improving performance are framed and discussed as (1) the mismatch between centralized animal husbandry and smallholder farming, (2) the mismatch between livestock density and cropland, (3) nutrient (including P) recycling and increasing organic matter content versus energy production, (4) subsidies for compost production and biogas, as well as (5) advances in the regulatory framework in China.     

Characterization of grain cadmium concentration in indica hybrid rice

As a consequence of contamination of soil with heavy metals, cadmium accumulation in grain is of great concern worldwide, but especially in southern China. It is important to evaluate the Cd accumulation potential of grain before or when examining and approving new cultivars. An evaluation method and criteria for verifying Cd accumulation potential in rice are proposed, and the Cd accumulation potential of 56 mid-season indica hybrids collected from the provincial cultivar trials in 2016 were investigated. Genotype, environment and their interactions strongly affected the variation in grain Cd accumulation. Two hybrids were identified as slightly Cd accumulating. Hybrids with slight Cd accumulation potential would be suitable for safe grain production on polluted land (total Cd under 2.0 mg·kg⁻¹) in Hunan Province (China) and should be considered for new cultivar evaluation and approval. This evaluation method and criterion could be applied for certifying Cd accumulation potential of rice cultivars.     

甘蓝型油菜花茎高效再生体系研究

研究了甘蓝型油菜花茎外植体高效诱导不定芽的再生体系,结果表明:以含有2,4-D(0.5￣1.0mg/L)的培养基对外植体进行预培养,以及在分化培养基中加入AgNO3(2￣6mg/L),可显著降低外植体褐化坏死的频率,提高了不定芽的发生频率及其再生能力;6BA(2.5mg/L)与NAA(0.1mg/L)配合使用有利于提高不定芽发生频率;再生的不定芽90%可长根。     

农杆菌介导大麦无筛选标记转基因植株的获得

【目的】目前,国内外大麦遗传转化主要利用Golden Promise品种,基因依赖性严重,尤其是大麦的转化效率较低,并且获得安全型转基因大麦植株对其进一步产业化非常重要。建立高效、无筛选标记大麦遗传转化体系,拓展大麦遗传转化的受体基因型,为大麦基因功能解析和大麦转基因育种及商业化种植提供技术保障。【方法】以优良大麦品种Vlamingh为受体,取开花授粉后14 d左右的幼胚为转化材料,通过对培养基成分及培养步骤优化,建立农杆菌介导的高效遗传转化体系,并利用该体系将Bar和GUS在不同T-DNA区段的双T-DNA表达载体pWMB123转化大麦,获得候选转基因植株,然后利用PCR、Bar试纸条、组织化学染色和Southern blot等检测方法,在T₁代转基因植株中成功获得无筛选标记大麦转基因植株。【结果】在愈伤组织分化阶段,发现培养基中添加1.0 mg·L^-1 KT、0.5 mg·L^-1 6-BA和0.05 mg·L^-1 NAA明显促进愈伤组织分化。在转基因植株生根阶段,发现采用添加1.0 mg·L^-1的IBA的SM1（无其他生长素）的生根效果最佳,培养基中添加2.5 mg·L^-1 CuSO₄显著降低了大麦转基因植株白化现象。共转化了138个幼胚,最终获得14株大麦转基因植株,转化效率10.14%。PCR、Bar试纸条、GUS染色等检测证实,T₀代转基因植株中均含有Bar,而仅有10株含有GUS,2个T-DNA的共转化效率为71.43%。选取4个同时含有Bar和GUS的转基因植株,对其自交后代进行检测,在BL8株系中筛选到2株只含GUS而不含Bar的转基因植株,无筛选标记效率为6.9%。在T₁代转基因植株中对Bar和GUS进行了Southern blot鉴定,发现在多数转基因植株中Bar和GUS均为多拷贝整合,进一步证实BL8-15和BL8-19为无筛选标记的转基因植株。【结论】利用大麦品种Vlamingh为转化材料可以较高效率获得转基因植株,提高愈伤组织分化效率和转基因植株生根效率,降低转基因植株白化现象。利用农杆菌介导双T-DNA表达载体转化大麦,成功获得了无筛选标记转基因植株。     

竹叶兰的无菌播种快繁技术研究

以竹叶兰种子为外植体进行无菌播种,成功诱导原球茎并再生植株,建立其快繁体系。试验结果表明：（1）1／2MS＋香蕉泥30．0g／L为最佳种子萌发与原球茎诱导培养基;（2）1／2MS＋6-BA1．0mg／L＋NAA0．1mg／L＋香蕉泥30．0g／L为最佳芽增殖培养基,且芽生长健壮;（3）1／2MS＋IBA0．5mg／L为最适生根培养基。     

百合遗传转化高频直接分化受体系统的构建

以东方百合西伯利亚和索蚌为试材,通过对鳞片分化成苗、小叶离体再生、生根和抗生素敏感性试验等研究,建立了稳定、高效的以无菌苗叶片为外植体的高频直接分化基因转移受体系统。在含有BA2mg/L+ZT1mg/L的MS培养基上,对供试品种无菌苗小叶片进行离体分化培养,再生率达100%,其适宜的生根培养基为MS+NAA0.75mg/L。同时,对小叶片外植体再生体系的抗生素及除草剂筛选浓度进行试验,结果显示:小叶片对G418和PPT较敏感,适宜的G418筛选浓度为80mg/L,PPT筛选浓度为1.75mg/L;而羧苄青霉素适宜浓度范围较宽,为300～400mg/L。     

Establishment of an efficient regeneration and genetic transformation system for Malus prunifolia Borkh. ‘Fupingqiuzi’

Malus prunifolia Borkh. ‘Fupingqiuzi’ has significant ecological and economic value and plays a key role in germplasm development and resistance research. However, its long juvenile phase and high heterozygosity are barriers to the identification of ‘Fupingqiuzi’ progeny with excellent traits. In-vitro regeneration techniques and Agrobacterium-mediated genetic transformation systems can efficiently produce complete plants and thus enable studies of gene function.However, optimal regeneration and...     

Oral administration of Allium sativum extract protects against infectious bursal disease in chickens

Garlic (Allium sativum, Liliaceae) has been safely used for more than 5000 years, and research on garlic extract is rapidly increasing because of its multiple biological functions. The in vivo effects of oral administration of garlic mixture (GM, water-soluble extract) on infectious bursal disease virus (IBDV)-infected specific pathogen free male white leghorn chicken were examined through histopathological, immunohistochemical, and Western blot analyses, and enzyme-linked immunosorbent assay. The results confirmed the protective effects of oral administration of 5 mg·kg⁻¹ BW GM (Group GM1) on bursal lesions after IBDV infection. In particular, protein expression of IBDV in the bursa decreased in Group GM1, indicating that GM administration decreased IBDV replication in the bursa. Furthermore, immunoglobulin M- and A-bearing B lymphocytes significantly increased 7 days post infection in bursae in Group GM1 (P<0.01), suggesting that the oral administration of 5 mg·kg⁻¹ GM offers moderate protection against B cell destruction after IBDV infection. During infection, the concentration of bursal interferon gamma (IFN-g) increased and peaked in Group GM1 earlier than in Group T (IBDV-exposed), demonstrating that GM administration prompted the production of IFN-g to protect against IBDV infection.     

12个网纹草品种的光合光响应曲线(简报)

测定了12个网纹草主要栽培品种的光合光响应曲线,通过曲线拟合和计算得到光饱和点、光补偿点等光合作用参数。结果表明:各品种最大光合速率均比较低,最高的是‘Red Star’,仅为4.74μmol·m^-2·s^-1;所有品种的光饱和点均在240～521μmol·m^-2·s^-1,因此在栽培时应控制光照条件,PAR小于520μmol·m^-2·s^-1为宜,否则易发生日灼;所有测试品种的光补偿点均小于7μmol·m^-2·s^-1,说明这些网纹草品种均可适应室内极弱光照条件。     

苦荞愈伤遗传转化体系的优化及用于FtCHS1的过表达分析

【目的】建立和优化苦荞愈伤组织遗传转化体系,为苦荞基因功能验证及分子育种提供研究工具。【方法】以苦荞品种“西荞二号”为材料,对苦荞愈伤遗传转化条件进行优化,包括苦荞外植体类型、诱导愈伤的激素比例、继代培养基的激素比例及农杆菌类型。利用苦荞类黄酮生物合成关键酶基因FtCHS1的过表达验证优化后的遗传转化体系。通过PCR筛选和荧光观察鉴定阳性株系,采用紫外分光光度法和高效液相色谱法（high performance liquid,HPLC）测定花青素及黄酮醇支路代谢物含量,使用实时荧光定量PCR分析类黄酮合成相关基因的表达,比较FtCHS1过表达愈伤组织与对照组的差异。【结果】苦荞诱导愈伤组织的最佳外植体为下胚轴,其最适诱导培养基为MS+0.8 mg·L^-1 6-BA+3.5 mg·L^-1 2,4-D,诱导率达72%;最优继代培养基为MS+3 mg·L^-1 6-BA+1 mg·L^-1 KT,愈伤组织增殖率与增殖系数分别为98%和1.09;转化过程中的最佳农杆菌是GV3101,转化效率达31.3%;FtCHS1过表达愈伤组织中,花青素、芦丁和杨梅素的含量显著高于对照（P<0.05）,山奈酚和槲皮素的含量极显著高于对照组（P<0.01）;外源FtCHS1的过表达对转基因愈伤组织中5个内源同源基因FtCHSs的表达水平没有影响（P>0.05）,而FtCHI、FtF3H、FtFLS1、FtFLS2、FtFLS3和FtDFR1等黄酮合成途径关键酶基因均上调表达（P<0.05）。此外,特异性正调控黄酮醇合成的转录因子基因FtMYB5和FtMYB6上调表达,而花青素合成抑制子基因FtMYB8的表达降低（P<0.05）。【结论】建立了苦荞愈伤组织遗传转化体系,过表达FtCHS1的苦荞愈伤组织通过上调黄酮合成相关基因的表达增加类黄酮物质的积累。     

K+ and Na+ fluxes in roots of two Chinese Iris populations

Maintenance of ion homeostasis, particularly the regulation of K⁺ and Na⁺ uptake, is important for all plants to adapt to salinity. Observations on ionic response to salinity and net fluxes of K⁺, Na⁺ in the root exhibited by plants during salt stress have highlighted the need for further investigation. The objectives of this study were to compare salt adaptation of two Chinese Iris (Iris lactea Pall. var. chinensis (Fisch.) Koidz.) populations, and to improve understanding of adaptation to salinity exhibited by plants. Plants used in this study were grown from seeds collected in the Xinjiang Uygur Autonomous Region (Xj) and Beijing Municipality (Bj), China. Hydroponically-grown seedlings of the two populations were supplied with nutrient solutions containing 0.1 (control) and 140 mmol·L^-1 NaCl. After 12 days, plants were harvested for determination of relative growth rate and K⁺, Na⁺ concentrations. Net fluxes of K⁺, Na⁺ from the apex and along the root axis to 10.8 mm were measured using non-invasive micro-test technique. With 140 mmol·L^-1 NaCl treatment, shoots for population Xj had larger relative growth rate and higher K⁺ concentration than shoots for population Bj. However, the Na⁺ concentrations in both shoots and roots were lower for Xj than those for Bj. There was a lower net efflux of K⁺ found in population Xj than by Bj in the mature zone (approximately 2.4-10.8 mm from root tip). However, no difference in the efflux of Na⁺ between the populations was obtained. Population Xj of I. lactea continued to grow normally under NaCl stress, and maintained a higher K⁺/Na⁺ ratio in the shoots. These traits, which were associated with lower K⁺ leakage, help population Xj adapt to saline environments.     

青州蜜桃、冬桃成熟胚子叶再生不定植株的研究

以青州蜜桃、冬桃成熟胚子叶为外植体 ,研究了利用子叶再生不定梢的适宜培养基。两个品种的子叶在 1 / 2MS附加BA 1 0mg/L和NAA 0 .2mg/L的培养基上获得了最高不定梢再生率。青州蜜桃和冬桃子叶的不定梢最高再生率分别是 6 6 .7%和 5 6 .2 %。不定梢在WPM +BA 2mg/L +IBA 0 .0 1mg/L +GA 0 .5mg/L培养基上增殖培养后 ,转移到 1 / 2MS +IBA 0 .5mg/L +蔗糖 2 0g/L诱导生根 ,两个品种的生根率都在 90 %以上     

柴蕉茎尖培养无菌株系建立与芽苗增殖

以柴蕉吸芽茎尖为外植体,进行无菌株系建立与芽苗增殖研究。试验结果表明,先将柴蕉吸芽接种于液体诱导培养基1/2 MS+4 mg.L-1BA+0.5 mg.L-1NAA+30 g.L-1蔗糖,7 d后转到1/2MS+4 mg.L-1BA+0.5 mg.L-1NAA+30 g.L-1蔗糖+1 mg.L-1AC中,可以有效减少外植体的褐变;在MS+4 mg.L-1BA+0.3 mg.L-1NAA+30 g.L-1蔗糖+6 g.L-1琼脂+80 g.L-1腺嘌呤培养基上采用薄切片进行继代增殖,出芽率高且芽苗粗壮。     

镉胁迫下黑曲霉TL-F2的促生特征及其对黑麦草种子萌发、幼苗生长和镉含量的影响

以黑曲霉(Aspergillus niger)TL-F2为供试菌株,黑麦草为供试植物,研究Cd胁迫下A. niger TL-F2的促生特性,以及对黑麦草种子萌发、幼苗生长和Cd含量的影响。结果表明:与不加Cd的对照组相比,低浓度Cd(5 mg·L^-1)胁迫下,A. niger TL-F2产吲哚乙酸(IAA)和溶磷的能力无显著变化,但产铁载体的能力显著(P<0.05)降低48.18%;中(20 mg·L^-1)、高(50 mg·L^-1)浓度Cd胁迫下,A. niger TL-F2产IAA的能力分别显著(P<0.05)降低55.76%和65.69%,溶磷量分别显著(P<0.05)降低50.07%和78.19%,产铁载体的能力分别显著(P<0.05)降低69.71%和80.08%。接种高浓度(1×10⁸ mL^-1)A. niger TL-F2有助于促进Cd胁迫下黑麦草种子的萌发和生长,高浓度Cd胁迫下,黑麦草地上部Cd含量较不接菌的显著(P<0.05)增加,增幅为17.95%;接种低浓度(1×10⁶ mL^-1)A. niger TL-F2对黑麦草种子萌发、Cd含量无明显促进作用。综合来看,接种适当量的A. niger TL-F2有助于促进Cd胁迫下黑麦草种子的萌发和生长,增加黑麦草地上部Cd含量,可作为黑麦草修复Cd污染水体和土壤的强化措施。