玉米浆发酵产生物丁醇的氨基酸代谢动力学模拟

为了深入挖掘利用丙酮丁醇梭菌产生物丁醇过程中氨基酸代谢的动态过程,探究利用廉价氮源玉米浆中的氨基酸用于丙酮丁醇梭菌产生物丁醇的生产策略,寻找生产丁醇的高效率廉价氮源来降低发酵生产成本。该研究首先利用高通量测序技术对玉米浆中微生物多样性进行分析;同时基于丙酮丁醇梭菌（Clostridium acetobutylicum）生产丙酮-丁醇-乙醇（Acetone-Butanol-Ethanol,ABE）碳代谢动态模型的基础上,构建氨基酸代谢模型,以此模拟15种氨基酸在利用木糖为碳源发酵生产ABE中的氨基酸代谢过程,并对氨基酸的代谢与丙酮丁醇梭菌的生物量以及ABE的合成相关性关系进行冗余分析;通过模型预测实际生产中利用玉米浆发酵时氨基酸的消耗过程。结果表明,梭状芽胞杆菌属（Clostridium）占细菌总数的68.76%,是玉米浆中的优势菌群;最佳参数校正后构建了氨基酸代谢模型,模拟值与试验值有较好拟合度;11种氨基酸（苯丙氨酸、苏氨酸、异亮氨酸、亮氨酸、蛋氨酸、缬氨酸、酪氨酸、甘氨酸、丝氨酸、精氨酸、天冬酰胺）在培养过程中迅速消耗用于细胞生长和溶剂生成,3种氨基酸（脯氨酸、组织胺、天冬氨酸）保持稳定状态,同时发酵过程中谷氨酰胺积累;冗余分析表明其中5种氨基酸对发酵产物及生物量影响具有相关性（P<0.05）,相关性排序从大到小依次为丝氨酸、甘氨酸、亮氨酸、缬氨酸、天冬酰胺;模拟预测玉米浆中缬氨酸、甘氨酸、丝氨酸在发酵过程中基本被消耗,推测其为发酵后期的营养限制性因子。该结论可证实玉米浆可作为丙酮丁醇梭菌发酵丁醇的优势氮源,为丙酮丁醇梭菌的氨基酸代谢调控及下一步利用并优化玉米浆作为氮源生产生物丁醇提供一定的理论参考和数据支撑。     

Effect of organic acids and nitrogen source on alcoholic fermentation: study of their buffering capacity

The effect of tartaric acid and other organic acids on alcoholic fermentation was studied. Organic acids in media with high sugar concentrations and ammonium as the sole nitrogen source had an enormous impact on Saccharomyces cerevisiae metabolism during alcoholic fermentation. The main effect on yeast metabolism was the quick acidification of the media in the absence of organic acids. All of the organic acids used in this study (tartaric, malic, citric, and succinic acids) showed a buffering capacity, but not all of the acids had the same one. However, the results suggested that buffering should not be considered the only effect of organic acids on yeast metabolism. Nitrogen source also had a great influence on media pH. Ammonium consumption by yeasts produced a greater acidification of the media than when amino acids were used.     

生物质热解过程中氮迁移转化机理研究进展

生物质热解产物中热解气和热解油具有较高能源利用价值,可作为替代燃料或化工原料,但伴随热解过程迁移至热解气/油中的氮元素不仅会影响其品质,热解气/油进一步利用后也会污染大气环境。该研究围绕生物质资源制备清洁能源的总目标,系统分析生物质热解过程中氮迁移转化机理,重点论述气相氮、液相氮和焦炭氮的生成与转化机理。通过总结前人研究,得出生物质热解气中的含氮物质主要为HCN、NH3等,其中NH3主要来源于氨基酸热解释放的氨基以及HCN在焦炭表面的水解转化;HCN主要来源于腈、含氮杂环等一次热解产物的二次裂解;热解油中的含氮物质主要为含氮杂环、腈与酰胺,其中含氮杂环主要由部分氨基酸片段或氨基酸间的脱水缩合反应产生;腈主要来源于氨基酸分子脱H2反应以及酰胺脱H2O反应;酰胺主要来源于NH3与羧基的置换反应。不同生物质种类与热解工况下氮的迁移转化特性复杂多样,生物质种类以及热解过程中的压力、停留时间、升温速率、温度、热解气氛、粒径、催化剂等因素均会影响热解过程中氮的迁移转化路径,最终影响生物质热解气/油中含氮物质的组成及分布。进一步提出生物质热解过程中氮排放控制未来研究方向,以期为实现农村生物质资源高效清洁利用提供参考。     

Use of nitrogen compounds in spontaneous and inoculated wine fermentations.

In this paper the use of nitrogen compounds in garnacha must inoculated with active dry wine yeast Saccharomyces cerevisiae subsp. cerevisiae strain Na33 has been studied. The results are compared to garnacha must fermented with indigenous yeasts (control must). In the samples where the inoculated yeast predominated, no qualitative differences were appreciated in the use of amino acids with respect to the control samples, although there were quantitative differences. In the musts where the Na33 strain dominated, a lesser quantity of amino acids were consumed at the beginning of fermentation than in the control samples. For that reason, probably, this yeast showed problems in competing for the nitrogen nutrients of the must; this could have made its implantation in one of the inoculated samples more difficult. At the end of fermentation the Na33 strain continued to consume amino acids at high concentrations of ethanol. Its high tolerance to this toxic could be favored by the production and rehydration of dry wine yeast in the presence of air.     

Corn Endosperm Fermentation Using Endogenous Amino Nitrogen Generated by a Fungal Protease

Fractionating the corn kernel to separate endosperm from germ and pericarp improves corn ethanol processing by increasing fermentation throughput and generating salable coproducts. One fractionation technology, dry fractionation (DF), suffers from loss of germ‐derived nutrients and amino acids, resulting in poor fermentation performance. Such deficiencies may be addressed by increasing nitrogen and other nutritional supplementation. As an alternative to exogenous nitrogen source, we investigated the use of a fungal protease to generate free amino nitrogen (FAN) from corn endosperm. Incubation of endosperm with protease did not affect subsequent liquefaction and saccharification. FAN supplementation through proteolysis resulted in fermentation being 99% complete in 48 hr, compared to 93% maximum with urea supplementation. Viable cell growth rates were similar in FAN and urea‐supplemented fermentations. Urea and FAN addition resulted in similar fermentation characteristics and similar FAN consumption rates as with FAN alone, which was indicative that FAN was assimilated preferentially. Increased amounts of maltose remaining after fermentation were correlated with initial FAN concentrations in mash. This observed trend was implicated in ethanol yield reduction of 2 g/L at high protease loading (generating 1.6 mg of FAN/g of glucose substrate) compared to a urea control. Using a glucose and maltose solution, we confirmed higher residual maltose in fermentations supplemented with high FAN concentrations. Use of protease to generate optimal FAN concentration in mash (1.2 mg of FAN/g of glucose substrate) could improve economics of dry fractionated corn ethanol production by increasing fermentation rates and, consequently, reducing fermentation time.     

Study of some Saccharomyces cerevisiae strains for winemaking after preadaptation at low temperatures

Low-temperature fermentations (13 degrees C) are considered to improve wine aromatic profiles. However, because the risk of stuck and sluggish fermentations is high, these fermentations are not common. The aim of this paper was to analyze the effect of different preadaptation protocols in two commercial wine strains on the fermentation and some wine parameters. Preadaptation is understood to be the process between the rehydration of active dry yeast and the inoculation. In this study, it consisted of preparing a fermentation starter (addition of yeast grown at 25 degrees C) or inocula preadapted at low temperatures (as before, but grown at a fermentation temperature of 13 or 17 degrees C). These results were compared with those of rehydrated active dry yeast, and a commercial "cryotolerant" yeast was used as a reference. General fermentation kinetic parameters, yeast imposition, nitrogen consumption, and main wine products were analyzed. The results showed that the preadaptation of a yeast could improve the fermentation performance, although this improvement was strain-dependent. Low-temperature fermentations also had some general effects: reduction of acetic acid and fusel alcohol production and increased concentrations of glycerol. When the yeast performed better in fermentation because of preadaptation, nitrogen consumption was faster and the wine's "negative" attributes (acetic acid, fusel alcohols) were significantly reduced. Thus, in some strains, preadaptation could be an effective mechanism for improving low-temperature fermentation, which also significantly reduces detrimental wine attributes.     

Effect of increased cobalt treatments on sewage sludge amended soil: Nitrogen species in soil and tranference to tomato plants

The effects of increased cobalt additions (0, 50, 100 and 200 v mg v kg m 1 soil) in sewage sludge-amended soil on organic matter, N Kjeldahl, ammonium and nitrate were studied in this experiment. Three different rates of sewage sludge were applied (0, 30 and 60 v tn v ha m 1 ) to soil as main plots, using tomato (Lycopersicum esculentum Mill var. Ramy) such testing cultivation. Plant biomass and nitrogen content in tomato leaf were also monitored. The organic matter in the soil was clearly affected by the fertilization. N Kjeldahl, ammonium and nitrate were favoured by organic treatments. Co seemed to reduce the transformation of ammonium to nitrate on amended soils, with accumulation of ammonium forms, especially at the higher application rates of sewage sludge. This incidence of Co on nitrogen species in soil decreased with the time of experiment, probably due to the reduction of availability of the pollutant. Aerial biomass production and nitrogen content in leaf were increased for the organic fertilization compared to the control. Only very high levels of Co in soil reduced significantly the aerial biomass production of tomato plants in amended soils. Co seemed to induce a decrease of the nitrogen in leaf in the amended soils, but not for the non-fertilized soils.     

Provision of carbon skeletons for amide synthesis in non-nodulated soybean and pea roots in response to the source of nitrogen supply

Abstract

Soluble amino acids in roots and primary amino acids, which were involved in primary ammonium assimilation, in the metabolites of ¹⁴C-glucose fed to roots for 3 h in the dark were analyzed in the roots of non-nodulated soybean and pea plants grown in ammonium, nitrate or nitrogen-free media for 1 day. Compared with the effect of nitrate, ammonium supply strongly affected the content and synthesis of the amino acids in the roots. In both soybean and pea roots, the supply of ammonium increased considerably the concentrations of the primary amino acids, and asparagine was the most predominant amide, followed by glutamine. In nitrate-supplied soybean roots, the concentrations of asparagine, aspartate and alanine increased, but the concentration of glutamine was low. In the roots of pea plants grown in nitrate media, asparagine was the predominant amino acid, although the composition of the primary amino acids was little affected by nitrate supply. The proportion of amino acids synthesized from ¹⁴C-glucose increased and asparagine rather than glutamine was predominantly synthesized in ammonium-supplied soybean and pea roots, whereas in nitrate-supplied roots asparagine was more actively synthesized than glutamine, although asparagine was not predominant. The ratio of C4 (asparagine + aspartate) to C5 (glutamine + glutamate) amino acids was twofold higher in ammonium-supplied and nitrate-supplied soybean roots than in roots receiving no nitrogen. In contrast, in pea roots, the C4/C5 ratio was twofold higher only in ammonium nutrition. The results obtained suggest that the roots of leguminous plants might possess an indigenous ability to provide a carbon skeleton for preferential synthesis of asparagine rather than glutamine with a high intensity of ammonium supply.     

不同干湿交替灌溉与氮肥形态耦合下水稻根系生长及功能差异

【目的】适宜的灌溉方式及氮肥管理是水稻高产高效的有效途径,大多数研究集中于地上部分及产量品质的形成,而对于根系形态生理及其与产量之间的关系研究还不够深入。本文探讨了干湿交替灌溉方式与氮肥形态耦合对水稻根系形态、生理及代谢的影响,探索干湿交替灌溉与氮肥形态耦合机理,为水稻高产及根系生理提供理论依据。【方法】试验于2016和2017年在河南科技大学试验农场进行,以徐稻3号为材料,供试土壤土质为黏壤土,采用灌水方式和氮肥形态二因素随机试验,设置CK [浅水层灌溉(0 kPa)]、WMD[轻度干湿交替灌溉(–20 kPa)]和WSD[重度干湿交替灌溉(–40 kPa)] 3种灌溉方式。氮肥供应设置铵态氮∶硝态氮三个混合比例处理:100∶0、50∶50 (即1∶1)、0∶100,由硫酸铵、硝酸铵和硝酸钠提供氮源。在分蘖盛期、穗分化始期、抽穗期和成熟期采样,以水稻茎基部为中心,挖取20 cm (长)×20 cm (宽)×30 cm (深)的土块测定干湿交替灌溉和氮肥形态处理的水稻根长、直径、表面积、体积、根尖数等根系形态指标、根系氧化力,采集根系伤流液分析其中氨基酸、蛋白质、可溶性糖含量以及在成熟期测定产量和产量构成。【结果】灌溉方式与氮肥形态之间存在显著的互作关系。WMD与铵硝1∶1耦合后水稻产量最高,达到1015.8 g/m^2,为本试验的最佳互作组合模式。WMD下,铵硝1∶1处理主要生育时期的根长显著增加了10.6%~17.0%,平均根直径增加了3.98%~25.25%,根体积增加了5.27%~26.40%,根表面积增加了6.27%~25.19%,提高了根尖数、根系伤流液中氨基酸、蛋白质、可溶性糖的含量,促进了根系的碳氮代谢和对养分、水分的吸收。WSD降低单位面积穗数及每穗粒数,显著降低水稻产量,铵硝100∶0处理平均降低38.20%、铵硝1∶1平均降低29.94%、铵硝0∶100平均降低35.0%,减少了根系长度,降低根体积、根表面积、根尖数,抑制根系活力及伤流液中物质的合成,不利于根系功能的维持。不同水分条件下氮肥形态对根系的影响不一,CK下,100%NH4^+处理根长及根系活力提高,而在WMD下,硝铵1∶1处理改善根系形态、提高根系活性,促进根系碳氮代谢,100%NO3^–处理不利于根系生长及根系功能的维持。水稻根长、根体积、根表面积、根尖数、根系活力与产量呈显著或极显著的正相关关系。施用100%NO3^–处理单位面积穗数下降,产量降低明显。【结论】轻度适宜的干湿交替灌溉配合施用一定比例的铵硝混合氮肥可以充分发挥水肥的耦合效应,促进强健根系形态的建成,提高根系的碳氮代谢及养分吸收利用,从而促进水稻的高产稳产。     

Digestion residues of typical and high-beta-glucan oat flours provide substrates for in vitro fermentation

In vitro fermentabilities of the oat flour digestion residues (ODR) from two commercial oat lines with 4.7 and 5.3% beta-glucan and from two high-beta-glucan experimental lines with 7.6 and 8.1% beta-glucan were evaluated and compared with fermentations of lactulose, purified oat beta-glucan (POBG), and purified oat starch (POS). Substrates were fermented by using an in vitro batch fermentation system under anaerobic conditions for 24 h. The progress of the fermentation was studied by following the change in pH of the fermentation medium, production of short-chain fatty acids (SCFA) and gases, and consumption of carbohydrates. The substrate from the flour with the greatest amount of beta-glucan tended to have the greatest pH decline and the greatest total SCFA production. A significant correlation occurred between gas production and SCFA formation (R 2 = 0.89-0.99). Acetate was produced in the greatest amounts by all of the substrates except POBG, by which butyrate was produced in the greatest amount. More propionate and butyrate, but less acetate, were produced from high-beta-glucan ODR. With the given fermentation conditions, >80% of the total carbohydrate was depleted by the bacteria after 24 h. Glucose was the most rapidly consumed carbohydrate among other available monosaccharides in the fermentation medium. Overall, the high-beta-glucan experimental lines provided the best conditions for optimal in vitro gut fermentations.     

Changes in nitrogen compounds in must and wine during fermentation and biological aging by flor yeasts.

Urea, ammonium, and free amino acid contents were quantified in a must from Vitis vinifera cv. Pedro Ximenez grapes and in fermented wine and after a short aging of this wine by Saccharomyces cerevisiae race capensis yeast under variable oxygen availability conditions. The previous compounds were also determined in a wine in which the nitrogen source was depleted by the same race of flor yeast (old wine) and also following the addition of ammonium ion, L-glutamic acid, and L-proline. Under specific conditions such as low oxygen level and the absence of some nutrients, the yeasts release some amino acids including L-threonine, L-tryptophan, L-cysteine, and L-methionine to the medium. These amino acids must originate primarily in a de novo synthesis from ethanol that regenerates NAD(P)+. On the basis of these results, the yeasts may be able to use amino acids not only as nitrogen sources but also as redox agents to balance the oxidation-reduction potential under conditions of restricted oxygen, when electron transport along the respiratory chain may be hindered or limited.     

Carbon skeletons for amide synthesis during ammonium nutrition in tomato and wheat roots

Heavy application of ammonium nitrogen to plant roots results in the conversion of ammonium nitrogen to the nitrogen of amides, glutamine, and asparagine, which are stored in roots or translocated to shoot. Since the net synthesis of such amides requires the supply of corresponding carbon skeletons, the carbon metabolism in amide synthesis in response to ammonium supply was investigated in tomato and wheat roots. The content of major primary amino acids was determined in tomato and wheat roots during a 4-d period of ammonium nutrition after 1-d culture in nitrogen-free nutrient solution. Ammonium supply led to a continuous increase in the asparagine content in wheat roots, whereas in tomato roots, the glutamine content increased 1 d after ammonium supply and thereafter the glutamine content was higher than the asparagine content. The amounts of amino acids synthesized from glucose-¹⁴C increased while the amounts of organic acids decreased in tomato roots by the supply of ammonium nitrogen for 1 d, compared to the roots that did not receive nitrogen. In tomato roots, the proportion of labeled glutamine was higher than that of labeled asparagine and the C5 amino acids were more strongly labeled than the C4 amino acids. These findings were different from the previous ones in wheat roots where the proportion of asparagine was found to be extremely high (Koga and Ikeda 2000: J. Fac. Agr. Kyushu Univ45, 7–13). To examine the in vivo asparagine synthesis, aspartate-'4C was fed to the roots. The labeling of asparagine, which was the most strongly labeled amino acid among the free amino acids, was remarkably strong in wheat roots whereas the labeling of glutamine was also pronounced in tomato roots. These results indicate that the ability to replenish carbon skeletons for amide synthesis in ammonium nutrition is different between tomato and wheat roots.     

氨基酸发酵尾液可促进樱桃番茄对水溶肥料氮素的吸收利用

【目的】 氨基酸发酵尾液因富含多种有机营养成分,近年来被作为增效材料在肥料生产领域广泛应用。本文利用15N标记技术,研究了氨基酸发酵尾液对水溶肥料氮素利用效果的影响。 【方法】 以樱桃番茄为供试作物进行盆栽试验。试验共设置8个肥料处理,分别为:焦磷酸钾和氯化钾为材料配制水溶肥料 (F,N–P₂O₅–K₂O比例为0–14–24);15N尿素 (15N丰度为10.46%)、焦磷酸钾和氯化钾为材料配制水溶肥料 (P,N–P₂O₅–K₂O比例为18–14–24);在两个肥料基础上分别添加谷氨酸发酵尾液 (H)、赖氨酸和苏氨酸混合尾液 (B) 和聚合谷氨酸尾液 (S) 配制水溶肥料FH、FB、FS、PH、PB和PS(发酵尾液添加量为200 g/L)。每个处理重复6次,随机区组排列。供试肥料于番茄定植、坐果、膨果、初果和盛果五个时期等量施入,其中,肥料P、PH、PB和PS按等尿素氮量施入,每次施入量为纯氮60 mg/kg风干土,肥料F、FH、FB和FS保持相同施入量。植株保留三穗果实打顶,收获后测量番茄株高、茎粗、生物量和产量,并对植株各部和收获后土壤中全氮含量和15N丰度进行测定。 【结果】 1) 水溶肥料添加氨基酸发酵尾液可以明显优化肥料氮素在促生、增产方面的作用效果。与P处理相比,PH、PB和PS处理的番茄株高、茎粗、地上部干重、根干重和果实产量分别平均增加17.7%、3.8%、8.5%、12.5%和4.8%。2) 水溶肥料添加氨基酸发酵尾液可以显著提高植株对肥料氮素的吸收量,增加叶片、果实等主要功能部位的肥料氮累积量,提高氮素利用率。较P处理,施用水溶肥料PH、PB和PS处理的番茄植株总氮吸收量、肥料氮吸收量分别平均增加8.1%和9.9%,叶片、果实等主要功能部位肥料氮累积量分别平均增加18.2%和8.0%,肥料氮素利用率平均提高9.9%。 【结论】 水溶肥料中添加氨基酸发酵尾液,可以显著促进作物生长,增加作物产量,促进作物对肥料氮的吸收累积,从而有效地提高作物对水溶肥料中氮素的吸收利用。      

Interactions of bacteria,protozoa and plants leading to mineralization of soil nitrogen

The capacity of bacteria and protozoa to mineralize soil nitrogen was studied in microcosms with sterilized soil with or without wheat plants. The effect of small additions of glucose or ammonium nitrate or both, twice a week was also tested. Plant dry weight and N-content, number of microorganisms and biomass plus inorganic N were determined after 6 weeks.The introduction of plants profoundly influenced the N transformations. In the presence of root-derived carbon, much more N was mineralized from the organic matter and immobilized mainly in plant biomass. “Total observable change in biomass N plus inorganic N” was negative in the unvegetated soils without additions, while a mineralization of 1.7 mg N microcosm^?1 was observed in microcosms with wheat plants grown with bacteria only. When protozoa were included, the N taken up by plants increased by 75%. Sugar additions resulted in an 18% increase of total N in the shoots when protozoa were present, but had no significant effect in the absence of grazers. Plants with the same root weight were more efficient in their uptake of inorganic N when protozoa were present. Plants grown with protozoa also had a lower R/S ratio, indicating a less stressed N availability situation. The lowest ratio was found with N additions in the presence of protozoa.The results indicate that, with energy supplied by plant roots or with external glucose additions, soil bacteria can mineralize N from the soil organic matter to support their own growth. Grazing of the bacteria is necessary to make bacterial biomass N available for plant uptake.     

氮素形态对解磷细菌发酵液解磷效果及小白菜生长和品质的影响

采用盆栽试验,研究了施用硝态氮、铵态氮对解磷细菌发酵液不同组分解磷活性及其对小白菜产量、品质的影响。结果表明,施用相同氮肥时,浇灌解磷细菌原发酵液处理 T3的作用效果明显优于菌体悬浮液T2及菌液上清T1;以硝态氮为底肥时,施用发酵原液的土壤有效态磷含量较高,小白菜产量最高,品质明显优化,即硝态氮含量下降, Vc、可溶性糖、可溶性蛋白、纤维素以及植株磷含量显著增加,且增加变幅均高于以铵态氮为底肥的效果。由此可知,硝态氮与解磷细菌发酵原液配施时, W1菌株解磷效果更好,可能是解磷菌株W1最适的氮源。因此,种植小白菜时直接向其根际施用解磷细菌发酵液并配施硝态氮肥,更有利于提高小白菜的品质及产量。     

Nitrogen assimilation in sunflower leaves and upward and downward transport of nitrogen

The assimilation of ammonium and nitrate nitrogen into amino acids of mature sunflower leaves and their transport to the other plant parts were investigated using nitrogen-15 as a tracer. In the leaf, to which ¹⁵N-labelled ammonium was vacuum-infiltrated, the ¹⁵N content of glutamine was always the highest of the amino acids tested and that of alanine was higher than that of glutamic acid and aspartic acid at 15 min after the infiltration. On the other hand in the leaf to which ¹⁵N-labelled nitrate was vacuum-infiltrated, the ¹⁵N content of glutamic acid and aspartic acid was superior to that of glutamine. The incorporation of ¹⁵N into serine was not active in the case of either ¹⁵N-labelled ammonium or nitrate. In the internodes above and below the treated leaf, through which photosynthates were transported into other parts, the ¹⁵N content of γ-aminobutyric acid and glutamine was markedly high when both nitrogen sources were supplied. There were no differences in the labelling patterns of amino acids between the upper and lower internodes. From these results it may be concluded that glutamine, glutamic acid, and aspartic acid play an important role in the assimilation of ammonium and nitrate nitrogen in leaves and that nitrogen is transported mainly in the forms of γ-aminobutyric acid and glutamine from the leaves to the other plant parts,     

叶面追施硝态氮肥抑制菠萝营养生长效应

采用叶面淋施的盆栽试验方法,以我国菠萝主栽品种-巴厘为试材,研究不同形态氮素对盆栽菠萝营养生长和菠萝叶片黄化的影响,为菠萝氮肥合理施用提供参考。试验结果表明,叶面淋施硝态氮处理的菠萝根、茎叶生物量显著低于叶面淋施铵态氮、酰胺态氮,黄叶数显著高于叶面淋施铵态氮、酰胺态氮。与铵态氮相比,硝态氮处理的菠萝总叶数、根数目、根重、茎叶重分别减少18.7%、26.5%、49.7%、43.5%,黄叶数增加192.7%。叶面淋施硝态氮抑制菠萝营养生长主要机理是硝态氮提高了土壤p H值,减少了铁吸收,降低菠萝叶片中全铁、活性铁、叶绿素含量(与铵态氮相比,分别减少25.9%、66.9%、23.2%)。     

Responses to nitrogen sources and regulatory properties of root phosphoenolpyruvate carboxylase

Time course of changes in extractable root phosphoenolpyruvate carboxylase (PEP C) activity was investigated in wheat, barley, and tomato plants fed with different nitrogen sources. Ammonium-fed plants exhibited a 2–2.5-fold higher PEPC activity than nitrate-fed plants at 7 d after the onset of nitrogen supply. Western blot analysis revealed that the amounts of PEPC subunit proteins increased gradually as reflected in the extractable PEPC activity. These results suggest that the increase in PEPC activity may be due to de novo protein synthesis. PEPC was SO-fold purified from tomato roots after several chromatographic steps. Metabolite effects on the partially purified enzyme were also investigated under optimal or suboptimal conditions in terms of pH and concentrations of phosphoenolpyruvate. Organic acids and acidic amino acids inhibited the enzyme activity, while hexose phosphates stimulated it. Glutamine and asparagine produced in the course of ammonium assimilation hardly affected the activity.     

Effects of nitrogen rate and nitrogen form on glycine uptake by pakchoi (Brassica chinensis L.) under sterile culture

It is well known that plants are capable of taking up intact amino acids. However, how the nitrogen (N) rates and N forms affect amino acid uptake and amino acid nutritional contribution for plant are still uncertain. Effects of the different proportions of nitrate (NO₃^?), ammonium (NH₄⁺) and ¹⁵N-labeled glycine on pakchoi seedlings glycine uptake were investigated for 21 days hydroponics under the aseptic media. Our results showed that plant biomass and glycine uptake was positively related to glycine rate. NO₃^? and NH₄⁺, the two antagonistic N forms, both significantly inhibited plant glycine uptake. Their interactions with glycine were also negatively related to glycine uptake and glycine nutritional contribution. Glycine nutritional contribution in the treatments with high glycine rate (13.4%–35.8%) was significantly higher than that with low glycine rate (2.2%–13.2%). The high nutritional contribution indicated amino acids can serve as an important N source for plant growth under the high organic and low inorganic N input ecosystem.     

Tannins in tropical browses: effects on in vitro microbial fermentation and microbial protein synthesis in media containing different amounts of nitrogen

Four species of browses (Acacia angustissima, Acacia salicina, Calliandra calothyrsus, andDichrostachys cinerea) were used to study the effect of tannins on microbial fermentation and microbial protein synthesis in incubation media containing high nitrogen (HN) and low nitrogen (LN) in the presence and absence of polyethylene glycol (PEG, MW 6000). The additional nitrogen in HN medium was supplied through ammonium bicarbonate. The use of HN medium significantly (P < 0.05) increased the in vitro gas and short-chain fatty acid (SCFA) production and microbial protein synthesis compared to the LN medium. Incubation of tannin-containing browses alone produced significantly (P < 0.05) lower gas and SCFA compared to in the presence of PEG in both HN and LN media. Inclusion of PEG in tannin-containing browses significantly (P < 0.05) reduced the molar proportion of propionate compared to in its absence. Higher N in the media resulted in 10.4 and 9.9% increases in in vitro gas and SCFA production, respectively, whereas inclusion of PEG to tannin-containing feed to remove the effect of tannins increased the in vitro gas and SCFA production by 186 and 195%, respectively, indicating that the low fermentation of tannin-containing browses could be due to the depressive effects of tannins on microbial activity and only partially accounted for by unavailability of N for rumen microbes. Incubation of browses with straw significantly (P < 0.05) decreased ammonia nitrogen concentration but increased the in vitro gas and SCFA production and microbial protein synthesis compared to straw alone.