某些环境条件对硝酸还原酶活性稳定因子的影响

小麦 Triticum aestivum L.苗在 NO_3~--N 完全营养液中培养比在 NH_4~ -N 完全营养液中培养,它们叶细胞内的硝酸还原酶(NR)即 NO_3-NR 比 NH_4-NR 活性增高了15倍,而它们叶片中的稳定因子(NR_(SF)),即 NO_3-NR_(SF)比 NH_4-NR_(SF)活化 NO_3-NR 的能力仅增加0.2倍,表明 NR 与 NR_(SF)不是依存关系;另外在 NO_3~--N 培养的黄化小麦叶片,及黄化缺氮、缺铝,加(?)的叶片中,所有的 NR_(SF)都十分稳定,并且保持较高活性,但这些叶片中没有测出NR 活性,因而认为,在植物叶细胞中,NR_(SF)不是调节 NR 活性的主要条件。     

Overexpression of Rice Genes OsNRT1.1A and OsNRT1.1B Restores Chlorate Uptake and NRT2.1/NAR2.1 Expression in Arabidopsis thaliana chl1-5 Mutant

Nitrogen uptake by plants is a key step for efficient nitrogen use, which affects plant growth and yield. Arabidopsis thaliana gene NRT1.1 was identified as a transporter related to nitrate (NO₃⁻) signaling and uptake. In rice, three orthologs of NRT1.1, named OsNRT1.1A, OsNRT1.1B, and OsNRT1.1C, have been identified. This study evaluated the potential of OsNRT1.1A, OsNRT1.1B, and OsNRT1.1C in NO₃⁻ signaling and uptake through overexpression in the Arabidopsis chl1-5 mutant. The expression of OsNRT1.1A, OsNRT1.1B, and OsNRT1.1C was evaluated in the roots and shoots of rice cultivated with NO₃⁻ or NH₄⁺. OsNRT1.1A was expressed in the roots and shoots cultivated with NO₃⁻ and NH₄⁺. OsNRT1.1B was expressed predominantly in roots of rice cultivated with NO₃⁻, while the expression of OsNRT1.1C was low in roots and shoots. Arabidopsis chl1-5 plants were transformed by the floral dip method using Agrobacterium tumefaciens to overexpress OsNRT1.1A and the alternative splicing product named OsNRT1.1As, OsNRT1.1B, and OsNRT1.1C. The chlorate test showed the ability of OsNRT1.1A, OsNRT1.1B or OsNRT1.1C to take up chlorate, as evidenced by the decrease in fresh weight. The OsNRT1.1B lineages presented higher toxicity to chlorate. Gene expression analyses showed that the insertion of OsNRT1.1A and OsNRT1.1B into Arabidopsis chl1-5 induced the expression of NRT2.1 and NAR2.1. OsNRT1.1As overexpression did not significantly affect the expression of NRT2.1 and NAR2.1. The results show the differential ability of NRT1.1 orthologs in rice to take up chlorate and signal the expression of other nitrate transporters, which may affect the efficiency of nitrogen utilization and its uptake.

     

Agronomic nitrogen‐use efficiency of rice can be increased by driving OsNRT2.1 expression with the OsNAR2.1 promoter

The importance of the nitrate () transporter for yield and nitrogen‐use efficiency (NUE) in rice was previously demonstrated using map‐based cloning. In this study, we enhanced the expression of the OsNRT2.1 gene, which encodes a high‐affinity transporter, using a ubiquitin (Ubi) promoter and the ‐inducible promoter of the OsNAR2.1 gene to drive OsNRT2.1 expression in transgenic rice plants. Transgenic lines expressing pUbi:OsNRT2.1 or pOsNAR2.1:OsNRT2.1 constructs exhibited the increased total biomass including yields of approximately 21% and 38% compared with wild‐type (WT) plants. The agricultural NUE (ANUE) of the pUbi:OsNRT2.1 lines decreased to 83% of that of the WT plants, while the ANUE of the pOsNAR2.1:OsNRT2.1 lines increased to 128% of that of the WT plants. The dry matter transfer into grain decreased by 68% in the pUbi:OsNRT2.1 lines and increased by 46% in the pOsNAR2.1:OsNRT2.1 lines relative to the WT. The expression of OsNRT2.1 in shoot and grain showed that Ubi enhanced OsNRT2.1 expression by 7.5‐fold averagely and OsNAR2.1 promoters increased by about 80% higher than the WT. Interestingly, we found that the OsNAR2.1 was expressed higher in all the organs of pUbi:OsNRT2.1 lines; however, for pOsNAR2.1:OsNRT2.1 lines, OsNAR2.1 expression was only increased in root, leaf sheaths and internodes. We show that increased expression of OsNRT2.1, especially driven by OsNAR2.1 promoter, can improve the yield and NUE in rice.     

杭州稻麦菜轮作地区大气氮湿沉降

通过雨水中NH+4-N/NO-3-N比率和铵态氮自然丰度值(δ15NH+4)的变化探讨大气氮湿沉降与农作施肥活动的关联性. 2003年6月至2005年7月,采用自行设计定制的雨水收集器在浙江杭州稻-麦-蔬菜轮作地区进行了为期2a的全天候连续雨水采样分析.结果显示,杭州稻-麦-蔬菜轮作地区雨水中NH+4-N/NO-3-N比率和δ15NH+4值呈现相似的季节性变化,雨水中NH+4-N/NO-3-N的峰值出现在6月底～9月上旬,而后逐渐下降,秋冬季(10～11月份)降到最低;来春麦菜集中施肥期(2～3月份),又呈现多个小高峰;5～6月份为单季稻和蔬菜基肥和追肥集中施用期,故而峰值也达4以上;入冬后仅在麦田施肥期出现一个小高峰而后明显下降,大都降到1以下;频繁施肥期雨水中的NH+4-N/NO-3-N比率值是农闲期的2～4倍,显示雨水NH+4-N/NO-3-N比率与农田施肥活动有密切关联与同步性,但与气温无直接关联(R2=0.0129).雨水中δ15NH+4值的变化,与雨水中NH+4-N/NO-3-N比率相似,呈现明显的季节性变化:稻麦生育后期与种前空闲期为正值,麦稻蔬菜集中施肥期转为负值.雨水中NH+4-N/NO-3-N比率与δ15NH+4值对大气湿沉降中氮的来源、形态及地面NH3排放源的强度有一定的表征意义.     

Cloning and functional characterization of a constitutively expressed nitrate transporter gene, OsNRT1, from rice

Elucidating how rice (Oryza sativa) takes up nitrate at the molecular level could help improve the low recovery rate (<50%) of nitrogen fertilizer in rice paddies. As a first step toward that goal, we have cloned a nitrate transporter gene from rice called OsNRT1. OsNRT1 is a new member of a growing transporter family called PTR, which consists not only of nitrate transporters from higher plants that are homologs of the Arabidopsis CHL1 (AtNRT1) protein, but also peptide transporters from a wide variety of genera including animals, plants, fungi, and bacteria. However, despite the fact that OsNRT1 shares a higher degree of sequence identity with the two peptide transporters from plants (approximately 50%) than with the nitrate transporters (approximately 40%) of the PTR family, no peptide transport activity was observed when OsNRT1 was expressed in either Xenopus oocytes or yeast. Furthermore, contrasting the dual-affinity nitrate transport activity of CHL1, OsNRT1 displayed only low-affinity nitrate transport activity in Xenopus oocytes, with a K(m) value of approximately 9 mM. Northern-blot and in situ hybridization analysis indicated that OsNRT1 is constitutively expressed in the most external layer of the root, epidermis and root hair. These data strongly indicate that OsNRT1 encodes a constitutive component of a low-affinity nitrate uptake system for rice.     

不同植物构成的人工湿地对生活污水中氮的去除效应

测定由不同植物构成的人工湿地的氨态氮、硝态氮和亚硝态氮含量,对比不同植物对生活污水中氮的去除效率.结果表明,与不种植物的人工湿地相比,由风车草[Cyperus alternifolius L. ssp. flabelliformis (Rottb.) Kiikenth.]、香根草[Vertiveria zizanioides (Linn.) Nash]、芦苇(Phragmitas communis Trin.)和美人蕉(Canna indica Linn.)构成的人工湿地对氨态氮去除率分别提高6%、8%、11%和14%;对硝态氮去除率分别提高5%、6%、13%和9%;对亚硝态氮去除率分别提高5%、7%、10%和7%,说明种植芦苇和美人蕉的人工湿地对生活污水中的氮具有较好的去除效果.     

不同生态稻区覆膜旱作稻氮营养生理及抗逆生理特性探讨

不同生态稻区生态环境和栽培管理方式不同,覆膜旱作稻氮营养生理和抗逆生理与常规水作稻有些异同,尤其是体内NO₃^--N和NH₄⁺-N含量及分布差异很大.杭嘉湖平原区覆膜旱作稻生长前期受干热天气影响,其分蘖期、拔节期、孕穗期体内NO₃^--N含量较水作稻有不同程度的降低,而NH₄⁺-N含量则极显著提高;金衢盆地覆膜旱作稻孕穗期体内NO₃^--N含量比常规水作稻高,根部NH₄⁺-N含量则显著降低,茎基部和叶片NH₄⁺-N含量有一定增加.正常气候条件下两生态稻区覆膜旱作稻孕穗期叶片硝酸还原酶(NR)活性和谷氨酰胺合成酶(GS)活性较水作稻均有不同程度的提高;丙二醛(MDA)、可溶性糖及脯氨酸(Pro)含量与水作稻相差不大,表明不同生态稻区的覆膜旱作稻可以通过不同的栽培管理模式,促进生长发育,达到高产目的.     

Homologous expression of γ-glutamylcysteine synthetase increases grain yield and tolerance of transgenic rice plants to environmental stresses

Various environmental stresses induce reactive oxygen species (ROS), causing deleterious effects on plant cells. Glutathione (GSH), a critical antioxidant, is used to combat ROS. GSH is produced by γ-glutamylcysteine synthetase (γ-ECS) and glutathione synthetase (GS). To evaluate the functional roles of the Oryza sativa L. Japonica cv. Ilmi ECS (OsECS) gene, we generated transgenic rice plants overexpressing OsECS under the control of an inducible promoter (Rab21). When grown under saline conditions (100 mM) for 4 weeks, 2-independent transgenic (TGR1 and TGR2) rice plants remained bright green in comparison to control wild-type (WT) rice plants. TGR1 and TGR2 rice plants also showed a higher GSH/GSSG ratio than did WT rice plants in the presence of 100 mM NaCl, which led to enhanced redox homeostasis. TGR1 and TGR2 rice plants also showed lower ion leakage and higher chlorophyll-fluorescence when exposed to 10 μM methyl viologen (MV). Furthermore, the TGR1 and TGR2 rice seeds had approximately 1.5-fold higher germination rates in the presence of 200 mM salt. Under paddy field conditions, OsECS-overexpression in transgenic rice plants increased rice grain yield (TGW) and improved biomass. Overall, our results show that OsECS overexpression in transgenic rice increases tolerance and germination rate in the presence of abiotic stress by improving redox homeostasis via an enhanced GSH pool. Our findings suggest that increases in grain yield by OsECS overexpression could improve crop yields under natural environmental conditions.     

缓/控释肥料研究进展

缓/控释肥料对作物产量的影响因作物种类、肥料种类和试验条件而异,大多数植物增产比较明显,如大豆使用长效尿素,与普通尿素相比增产幅度最高可达33%。各种类型的缓/控释肥料,均可不同程度的提高肥料利用率。缓/控释肥料在农业上的应用能有效地保护生态环境,如抑制土壤NH4 向NO3-氧化,减少土壤NO3-的积累,从而减少氮肥以NO3-形式淋溶损失,减少了施肥对环境的污染;可以减少土壤N2O的释放等。提出了目前缓/控释肥料在农业应用中存在的问题及今后的发展方向。     

pOsNAR2.1:OsNAR2.1 expression enhances nitrogen uptake efficiency and grain yield in transgenic rice plants

The nitrate () transporter has been selected as an important gene maker in the process of environmental adoption in rice cultivars. In this work, we transferred another native OsNAR2.1 promoter with driving OsNAR2.1 gene into rice plants. The transgenic lines with exogenous pOsNAR2.1:OsNAR2.1 constructs showed enhanced OsNAR2.1 expression level, compared with wild type (WT), and ¹⁵N influx in roots increased 21%–32% in response to 0.2 mm and 2.5 mm and 1.25 mm ¹⁵NH₄¹⁵NO₃. Under these three N conditions, the biomass of the pOsNAR2.1:OsNAR2.1 transgenic lines increased 143%, 129% and 51%, and total N content increased 161%, 242% and 69%, respectively, compared to WT. Furthermore in field experiments we found the grain yield, agricultural nitrogen use efficiency (ANUE), and dry matter transfer of pOsNAR2.1:OsNAR2.1 plants increased by about 21%, 22% and 21%, compared to WT. We also compared the phenotypes of pOsNAR2.1:OsNAR2.1 and pOsNAR2.1:OsNRT2.1 transgenic lines in the field, found that postanthesis N uptake differed significantly between them, and in comparison with the WT. Postanthesis N uptake (PANU) increased approximately 39% and 85%, in the pOsNAR2.1:OsNAR2.1 and pOsNAR2.1:OsNRT2.1 transgenic lines, respectively, possibly because OsNRT2.1 expression was less in the pOsNAR2.1:OsNAR2.1 lines than in the pOsNAR2.1:OsNRT2.1 lines during the late growth stage. These results show that rice NO₃^– uptake, yield and NUE were improved by increased OsNAR2.1 expression via its native promoter.     

葡萄糖存在下选择性抑菌剂对土壤氮素转化的影响

采用2种氮源并分别加入选择性微生物抑制剂进行室内培养,通过测定样品中NH₄⁺-N和NO₃^--N及土壤中氨基葡萄糖和胞壁酸含量,研究土壤微生物氮素固持时间特征及其相对贡献.结果表明,加入青链霉素明显地降低了NH₄⁺-N的转化速率,且影响远大于真菌抑制剂放线菌酮;氨基葡萄糖和胞壁酸的相对比例急剧增加,而后趋于平衡;加入放线菌酮后NO₃-N转化速率持续下降,氨基葡萄糖的合成受到抑制,但加入细菌抑制剂青链霉素对其转化无显著影响.培养初期,细菌在葡萄糖存在下能快速固持NH₄⁺-N和NO₃^--N,并以NH₄⁺-N为首选氮源;培养后期,氮转化主要为真菌所推动,且真菌对NO₃^--N的利用能力显著大于细菌.     

Gene structure and expression of the high-affinity nitrate transport system in rice roots

Rice has a preference for uptake of ammonium over nitrate and can use ammonium-N efficiently. Consequently, transporters mediating ammonium uptake have been extensively studied, but nitrate transporters have been largely ignored. Recently,some reports have shown that rice also has high capacity to acquire nitrate from growth medium, so understanding the nitrate transport system in rice roots is very important for improving N use efficiency in rice. The present study identified four putative NRT2 and two putative NAR2 genes that encode components of the high-affinity nitrate transport system (HATS) in the rice (Oryza sativa L. subsp, japonica cv. Nipponbare) genome. OsNRT2.1 and OsNRT2.2 share an identical coding region sequence, and their deduced proteins are closely related to those from monocotyledonous plants. The two NAR2 proteins are closely related to those from mono-cotyledonous plants as well. However, OsNRT2.3 and OsNRT2.4 are more closely related to Arabidopsis NRT2 proteins. Relative quantitative reverse tranecdption-polymerase chain reaction analysis showed that all of the six genes were rapidly upregulated and then downregulated in the roots of N-starved rice plants after they were re-supplied with 0.2 mM nitrate, but the response to nitrate differed among gene members.The results from phylogenetic tree, gene structure and expression analysis implied the divergent roles for the individual members of the rice NRT2 and NAR2 families. High-affinity nitrate influx rates associated with nitrate induction in rice roots were investigated and were found to be regulated by external pH. Compared with the nitrate influx rates at pH 6.5, alkaline pH (pH 8.0) inhibited nitrate Influx, and acidic pH (pH 5.0) enhanced the nitrate influx In I h nitrate induced roots, but did not significantly affect that in 4 to 8 h nitrate induced roots.     

草莓离体无性繁殖的研究——Ⅰ.影响草莓离体繁殖的因素

基本培养基对草莓的离体快速繁殖有重要影响。通过试验筛选出一个比目前常用的MS和White培养基分别高5.53倍和2.3倍的F培养基(总氮量15mM,其中硝态氮10mM,铵态氮5mM,其它成份同MS)。相同浓度的BA与不同的基本培养基配合作用效果相差悬殊。由于草莓茎尖在MS培养基上生长缓慢,本文探讨了在室温条件下进行草莓种质保存的可能性。还讨论了由花药培养获得再生植株在快速繁殖中的应用问题。     

多花黑麦草对啤酒废水净化功能的研究

1 引言 利用水生高等植物净化污水,国内外已有很多报道,1992年丁树荣等首先利用人工基质无土栽培多花黑麦草(Lolium multiflorum Lam.)净化缫丝废水获得成功,是利用陆生植物于高等水生植物净化塘中以净化有机污水的首例,并提出了解决在亚热带和温带冬季低温地区,利用高等水生植物净化污水时冬季不能有效运转难题的新途径,而作者于1987—     

一氧化氮对盐胁迫下小麦幼苗根生长和氧化损伤的影响

0.05和0.10 mmol/L一氧化氮(NO)供体硝普钠(sodium mtropmsside,SNP)处理明显减轻NaCl浓度为150 mmo1/L左右的盐胁迫对小麦幼苗根生长的抑制效应,其中0.05mmol/L的SNP效果最明显;0.30mmol/L以上的SNP处理对根抑制无明显缓解作用;当NaCl浓度大于300 mmol/L时,各种浓度的SNP均不能减轻盐胁迫对根生长的抑制.以N O清除剂血红蛋白(hemoglobin,Hb)以及NOx-,K3Fe(CN)6等为对照,观察到0.05 mmol/L的SNP能不同程度地提高150mmo/L盐胁迫下小麦幼苗根尖细胞中超氧化物歧化酶(SOD)、过氧化物酶(POD)和抗坏血酸过氧化物酶(ascorbateperoxidase,APX)活性,明显降低MDA、H2O2和O2-.的积累,阻断盐胁迫诱导的根尖细胞DNA片段化,表明NO能有效缓解盐胁迫引起的小麦幼苗根尖细胞的氧化损伤.     

氮素形态配比对菜用大豆籽粒膨大过程中氮碳同化的影响

以菜用大豆品种‘理想95-1'为试材,通过蛭石盆栽试验研究了氮素不同形态配比对菜用大豆籽粒膨大过程中碳氮代谢的影响.结果表明:营养液中增加适当的铵态氮比例(25%～50%)有利于菜用大豆的生长发育,菜用大豆植株和荚果干鲜重、干物率显著增加,尤以硝铵比为75∶25最为显著.在高比例的硝态氮(100%)或铵态氮(75%)处理下,菜用大豆籽粒的硝酸还原酶(NR)和谷氨酰胺合成酶(GS)活性分别显著升高,氮代谢显著增强,可溶性蛋白含量迅速增加,但同期的籽粒淀粉酶活性较低,可溶性糖和淀粉含量显著下降,碳代谢受到显著抑制.在硝铵比为75∶25时,菜用大豆籽粒氮代谢强度适中,同期碳代谢显著增强,籽粒可溶性糖和淀粉含量显著升高,能维持籽粒正常的生理代谢,有利于菜用大豆籽粒发育过程中营养物质的积累.可见,硝态氮和铵态氮配比能显著影响菜用大豆籽粒发育过程中的碳氮代谢,籽粒碳氮代谢与其产量密切相关,可通过调节硝铵比来获得理想菜用大豆产量.