IL15/PAP融合蛋白基因克隆及其原核与植物表达载体的构建

将IL15和PAP基因通过一甘氨酸接头(Gly4Ser)3连接在一起,构建原核和植物表达载体,以期表达具有导向杀伤活性的融合蛋白.在引物设计中,通过引物在IL15基因下游引入(Gly4Ser)3的编码序列和BamHI位点,然后采取分步克隆及PCR、酶切、连接等一系列分子克隆方法,将IL15和PAP基因融合在一起,构建融合蛋白的原核与植物表达载体.经PCR和NcoI/SalI双酶切鉴定及测序,证实原核表达载体pET32a-IL15/PAP及植物表达载体pB-I P中均插入了正确的融合基因序列.     

施用尿素、红萍对水稻生长及其氮素利用效率的影响

应用~(15)N示踪技术研究了尿素和红萍氮素对水稻分蘖、成穗和产量的影响以及当季水稻对氮素的利用率。结果表明:30十30kgN/ha尿素分别作基肥和分蘖期追肥(U-2),10十20十30kgN/ha尿素分别作基肥、分蘖肥和穗肥(U-3),30十30kgN/ha的红萍分别作基肥和分蘖期追肥(A-2),和不施氮肥对照(CK)处理,水稻分蘖总数分别为465.0,495.0,579.0,403.5万/ha;成穗率分别为81.9％,78.8％,69.8％,74.4％;每穗平均粒数分别为75.8,80.6,78.2,74.7粒。施红萍的稻谷产量与等氮量尿素相当。施用尿素和红萍处理,水稻生长前期平均出蘖速度明显不同,而且水稻不同时期分蘖,其成穗数和每穗粒数均随分蘖时间推移呈逐渐递减变化。试验结果表明:红萍作基肥当季的氮素利用率为41.3—48.2％,高于尿素作基肥的利用率(28.0—31.9％),但尿素作追肥(45.7—46.2％)则优于红萍作追肥(37.5—43.2％)。U-3,A-2,U-2处理当季氮素利用率分别为47.7—50.6％,39.2—45.7％,37.0—38.8％。     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

鲁棉研15号及其父母本核型对比分析

采用常规压片法对鲁棉研15号及其父本——转基因抗虫棉选系613、母本——常规陆地棉选系R55染色体数目和核型进行了研究。结果表明:鲁棉研15号及其父母本的染色体数目均为2n=26。鲁棉研15号的核型公式为:k(2n)=26=10m+12sm+4st,属于"2B"类型,染色体相对长度组成为2n=26=8M1+10M2+4L+4S。其父本的核型公式为:k(2n)=26=6m+2sm(SAT)+16sm+2st,属于"3B"类型,染色体相对长度组成为2n=26=8M1+8M2+4L+6S。其母本的核型公式为:k(2n)=26=20m+6sm,属于"2A"类型,染色体相对长度组成为2n=26=6M1+18M2+2S。通过比较发现,鲁棉研15号倾向于父本,三者中属母本最原始。     

Contribution of earthworm activity to the infiltration of nitrogen in a wheat agroecosystem

The effect of earthworms on leachate volume and leachate N losses was investigated in a rice–wheat rotation agroecosystem over the wheat growing season. Corn residues (<2 cm) were added to field microcosms as either mulch or incorporated into the soil. Earthworms were added to half of the microcosms at a population density and age structure simulating a natural earthworm population. With earthworms present, leachate volume was >30% higher, but this result was not statistically significant (P>0.05). Earthworms significantly increased mineral N losses in the mulching treatment (P<0.05). There was a reduced fertilizer N (urea-¹⁵N) loss when earthworms were present, but the reduction was not significant (P>0.05). Our results indicate that earthworm activity can accelerate leachate production and N loss to a certain extent under wheat, and that native soil N (including mineral N and organic N) and/or organic matter N (corn residue N) are more likely sources for N leaching than urea-N.     

小麦和玉米叶面标记尿素态15N的吸收和运输

以15N标记的尿素作为叶面施肥和根系后期追肥的N肥品种,分别采用土培与砂培研究小麦与玉米拔节后至灌浆初期不同生育期、不同N、P和K肥叶面配施后作物对叶面N肥利用效率、叶面施用N肥的分配及对全株N、P和K养分状况的影响。无论是小麦还是玉米叶片均能有效地吸收N素并将其迅速转移。玉米拔节期下位叶（第5叶）施肥,植株吸收的15N转移到根部的比例平均为9.0%,而中位叶（第8叶）处理,分配到根系的比例仅为2.4%。叶面施用尿素态氮肥的N在小麦地上部的回收率一般在54.5%～68.9%,在玉米全株体内的回收率可达59.9%～75.3%。随生育期推迟,两种作物叶片吸收N的回收利用率均明显提高。与单施氮肥相比,N,P和K肥配施均有提高氮素回收率和改善作物地上部N素营养的作用,尤以中位叶（第8叶）配施尿素＋KH2PO4的效果最佳,表明植物的叶面营养仍应注意平衡供应和适宜的施用时期与施肥部位。叶面大量元素营养不仅改善了所施肥料成分中相应元素的营养状况,同时也促进了对其他两种成分的吸收,并且提高了N、P转运到籽粒中去的比例。玉米吐丝期根系追施的尿素态N有56%积累在籽粒中,而叶面施肥所利用的N分配在籽粒中的比例为70%。     

红萍在稻田氮素平衡中的作用

应用１５Ｎ示踪法研究了红萍在稻田氮素平衡中的作用。红萍作基肥，当季水稻的１５Ｎ－红萍Ｎ素利用为２７．８％；第二季５．３８％；土壤残留３６．１％。红萍可排出体内１２％以上的氮素。红萍的排氮和吸氮有助于调节稻田的氮素平衡，稻田养萍抑制藻类生长，降低ｐＨ值和ＮＨ浓度，减少ＮＨ３挥发损失，以及减少蓝藻（颤藻）反硝化反应释放的Ｎ２Ｏ，提高化学肥料１５Ｎ的回收率：它比１５Ｎ－尿素不养萍处理１５Ｎ回收率增加４．２５％，比１５Ｎ－尿素追肥不养萍处理１５Ｎ回收率增加８．３５％～２５．１１％。     

北京地区潮土表层中NO3--N的转化积累及其淋洗损失

本试验利用渗滤池设施，采用化学分析和同位素技术相结合的方法研究了北京地区潮土表层中施用氮肥后ＮＯ３＾－－Ｎ转化积累及其１３０ｃｍ土体的淋洗状况。常规分析结果表明，在春小麦和夏玉米的生育前期可以观察到氮素明显地向ＮＯ３＾－－Ｎ的转化积累，其强度随尿素施用量的增加而明显增加，而尿素、硝铵、硫铵等不同氮肥品种处理之间有差异但大多不显著。同时夏玉米期间转化积累作用比春小麦期间强烈。＾１５Ｎ标记试验结果表明     

设施栽培油桃对叶面施15N的吸收、分配特性研究

以设施栽培的5年生早红珠油桃/山毛桃为试材,应用15N示踪技术研究油桃叶片对15N-尿素的吸收及运转特性。结果表明,叶片施用15N-尿素标记叶吸收主要发生在叶片涂抹15N-尿素后6.h内,平均吸收速率为0.204mg/(g.h);标记叶中15N吸收量24.h达到最高,新梢和梢顶嫩叶15N含量在施用15N-尿素48.h达到最高,下部叶15N含量很低,没有明显的峰;处理168.h各器官中15N含量为标记叶梢顶嫩叶新梢下部叶;试验结束时分配势Ndff(即各器官N含量来自化肥N所占的百分率)为标记叶中最高,然后依次为梢顶嫩叶、新梢、下部叶。这说明氮素迅速被吸收并运到嫩梢和嫩叶中,从而促进这些新生器官的形态建造,可起到以N增C的作用。不同叶面处理的试验还表明,正面和背面全部涂抹的叶片15N含量最高,只涂抹叶片背面次之,涂抹正面最低。设施栽培油桃叶片可迅速吸收尿素,其吸收量早晨明显优于中午和下午,因此设施油桃栽培管理中于早晨进行叶面施尿素,且正反面兼顾,N素的吸收利用效果最好。     

Release of C and N from roots of peas and oats and their availability to soil microorganisms

Nutrient mobilisation in the rhizosphere is driven by soil microorganisms and controlled by the release of available C compounds from roots. It is not known how the quality of release influences this process in situ. Therefore, the present study was conducted to investigate the amount and turnover of rhizodeposition, in this study defined as root-derived C or N present in the soil after removal of roots and root fragments, released at different growth stages of peas (Pisum sativum L.) and oats (Avena sativa L.). Plants were grown in soil columns placed in a raised bed under outdoor conditions and simultaneously pulse labelled in situ with a ¹³C-glucose-¹⁵N-urea solution using a stem feeding method. After harvest, ¹³C and ¹⁵N was recovered in plant parts and soil pools, including the microbial biomass. Net rhizodeposition of C and N as a percentage of total plant C and N was higher in peas than in oats. Moreover, the C-to-N ratio of the rhizodeposits was lower in peas, and a higher proportion of the microbial biomass and inorganic N was derived from rhizodeposition. These results suggest a positive plant-soil feedback shaping nutrient mobilisation. This process is driven by the C and N supply of roots, which has a higher availability in peas than in oats.