Purification and Trypsin Inhibitor Activity of a Sporamin B from Sweet Potato （Ipomoea batatas Lam. 55-2）

Sporamin is a soluble protein in sweet potato, and falls into two distinct homology groups, subfamilies A and B. In this research, a sporamin B was purified and its amino acid sequences, trypsin inhibitor activity （Ti activity） were analyzed. This sporamin B was isolated from sweet potato tubers [Ipomoea batatas （L.） Lam cv. 55-2] through extraction of the water-soluble fraction, dialysis, ultrafiltration and ion-exchange chromatography. Homology determined by polyacrylamide gel electrophoresis showed that mainly one bond appeared in gel after being reduced by SDS （sodium dodecyl sulfate）, or by SDS and 2-mercaptoethanol, or in native situation. By comparing the data of the polypeptide mass Matrix-assisted laser desorption/ionization time-of-flight （MALDI-TOF） mass spectrometry with those of the mass of the theoretical amino acid sequences from NCBI protein database, it was revealed that it was Q40091｜Q40091_IPOBA, sweet potato sporamin B - Ipomoea Batatas （sweet potato） （Batate）. The sequence coverage was 70.6%. N-terminal sequence was SETPV （Ser-Glu-Thr-Pro-Val）. There is a linear relationship between trypsin inhibitor activity （Ti activity） and amounts of this sporamin B （3-18 μg mL-1）. The equation of linear regression was y = 2.5809x ＋ 17.049 （r2 = 0.9966）. There was a curvilinear relationship between Ti activity and amounts of this sporamin B （21-150 μg mL-1）. The equation of curvilinear regression is y = 14.417ln（x） ＋ 23.26 （r2 = 0.9924）. The concentration of sporamin B with Ti activity after heating at 40°C may induce part denature of this sporamin B, and there was no statistic difference after heating at 40, 50, 60°C for 20 min. Heat treatment at more than 90°C leads to a dramatic decrease of trypsin inhibitor efficiency. The results suggested that Q40091｜Q40091_IPOBA was the major sporamin B in sweet potato tubers [Ipomoea Batatas （L.） Lam cv. 55-2], which had strong Ti activity, and was stable to both thermal and DTT （DL-dithiothreitol） relatively.     

灭幼脲类杀虫剂防治森林害虫的研究进展

本文论述了灭幼脲类杀虫剂目前的种类,杀虫机理,对人、畜等的安全性,害虫产生抗药性情况及其防治对象和防治方法的研究进展。最后提出灭幼脲这种昆虫生长抑制剂防治森林害虫,具有广谱、高效、低毒、低成本和残效期长等优点,而且生态效益、社会效益和经济效益明显,具有广阔的发展前途,目前已进入实际应用阶段。     

纤维素酶抑制剂对松墨天牛消化生理的影响

以松墨天牛的消化生理为基础，研究了松墨天牛消化酶中纤维素酶的动力学特性；以现代酶学研究方法为主，结合生物测定技术，进一步研究了不同性质和不同浓度的酶抑制剂对松墨天牛成虫离体和活体纤维素酶酶活力的影响，结果表明，选用的酶抑制剂无论对分离提取的纤维素酶液和活体中的纤维素酶8都具有显著的抑制作用。饲养试验中，经0.1%酶抑制剂处理的饲料组5d死亡率为75%.ck组平均成活天数为28d。     

Screening of extracts of Japanese woods for melanin biosynthesis inhibition

The effects of 120 methanol extracts prepared from bark and heartwood of 69 types of Japanese wood on the melanin production of B16 melanoma cells were examined. The melanin content of B16 melanoma cells was determined spectrophotometrically at 405nm. The extracts were also examined for their effects on cell viability. We found that the methanol extracts of Fagus crenata (buna, wood, 100μg/ml), Sapium sebiferum (Nankinhaze, wood, bark, 10μg/ml), and Zelkova serrata (keyaki, wood, 10μg/ml) greatly inhibited the melanin production of B16 melanoma cells without significant cytotoxicity. However, these extracts did not inhibit tyrosinase activity at the concentration of 100μg/ml. These findings indicate that the depigmenting mechanism of these extracts involves the suppression of some pigmenting signals in stimulating melanogenesis rather than the inhibition of tyrosinase activity. Part of this study was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, Japan, March 2003     

Hypoxia inducible factor 1α expression and effects of its inhibitors in canine lymphoma

Hypoxic conditions in various cancers are believed to relate with their malignancy, and hypoxia inducible factor-1α (HIF-1α) has been shown to be a major regulator of the response to low oxygen. In this study, we examined HIF-1α expression in canine lymphoma using cell lines and clinical samples and found that these cells expressed HIF-1α. Moreover, the HIF-1α inhibitors, echinomycin, YC-1 and 2-methoxyestradiol, suppressed the proliferation of canine lymphoma cell lines. In a xenograft model using NOD/scid mice, echinomycin treatment resulted in a dose-dependent regression of the tumor. Our results suggest that HIF-1α contributes to the proliferation and/or survival of canine lymphoma cells. Therefore, HIF-1α inhibitors may be potential agents to treat canine lymphoma.     

三种新型化合物对草地贪夜蛾海藻糖与几丁质代谢及生长发育的影响

[目的]几丁质是昆虫外骨骼和围食膜的主要成分,其合成始于海藻糖酶(trehalase,TRE),终于几丁质合成酶(chitin synthase,CHS),蜕皮与外表皮重塑过程则需要依靠几丁质酶(chitinase,CHT)完成.本研究通过注射3种新型化合物,检测草地贪夜蛾(Spodoptera frugiperda)...     

Effects of the nitrification inhibitor dicyandiamide on soil mineral N, pasture yield, nutrient uptake and pasture quality in a grazed pasture system

Recent lysimeter studies have demonstrated that the nitrification inhibitor, dicyandiamide (DCD), can reduce nitrate (NO) leaching losses from cow urine patches in grazed pasture systems. The objective of this study was to quantify the effects of fine particle suspension (FPS) DCD on soil mineral N components, pasture yield, nutrient uptake and pasture quality under grazed pasture conditions. A field study was conducted on the Lincoln University dairy farm, Canterbury, New Zealand, from 2002 to 2006. FPS DCD was applied to grazed pasture plots at 10 kg ha^?1 in early May in addition to applied cow urine patches at a nitrogen (N) loading rate of 1000 kg N ha^?1, with DCD reapplied in early August. Soil mineral N levels in the urine patches were monitored. Pasture yield, N and cation concentrations and uptake were measured in treatment urine patches and inter‐urine areas of the pasture. Comparisons were made with control plots which did not receive DCD. NO levels under the DCD‐treated urine patches (0–7.5 cm) were in the order of 10 kg N ha^?1 compared with 40–80 kg N ha^?1 under untreated patches, and soil ammonium (NH) levels were consistently higher under the DCD‐treated patches. The DCD significantly and consistently increased pasture yield in both the urine patches, and inter‐urine areas of the pasture in all 4 years of the trial. Mean annual dry matter (DM) yields over 4 years were inter‐urine areas, 10.3; inter‐urine + DCD, 12.4; urine, 12.4 and urine +DCD 16.0 t DM ha^?1, representing an average DM yield increase of 20 and 29% in inter‐urine and urine patch areas, respectively. On a whole paddock basis, the increase in annual DM yield resulting from DCD application was estimated to be 21%. N, calcium (Ca), magnesium (Mg) and potassium (K) concentrations in pasture were unaffected by treatment with DCD; however, total annual uptake of these nutrients by pasture was significantly higher in all years where DCD had been applied. Pasture DM, protein, carbohydrate, metabolizable energy and fibre levels and sward clover content were not affected by treatment with DCD. The results demonstrate the agronomic value of the DCD treatment in addition to the environmental benefits in a grazed pasture system.     

Cross-resistance profile of mesosulfuron-methyl-resistant Italian ryegrass in the southern United States

Diclofop-resistant Lolium species (ryegrass) is a major weed problem in wheat production worldwide. This study was conducted to determine the resistance pattern of diclofop-resistant ryegrass accessions from the southern United States to mesosulfuron-methyl, a recently commercialized herbicide for ryegrass control in wheat; to determine the cross-resistance pattern of a Lolium multiflorum Lam. (Italian ryegrass) accession, 03-1, to acetolactate synthase (ALS) and acetyl-CoA carboxylase (ACCase) inhibitors; and to determine the resistance mechanism of Italian ryegrass to mesosulfuron-methyl. Seventeen ryegrass accessions from Arkansas and Louisiana, including standard resistant and susceptible accessions, were used in this experiment. Fourteen of the 17 accessions were more resistant (four- to > 308-fold) to diclofop than the standard susceptible biotype. One accession, 03-1, was resistant to mesosulfuron-methyl as well as to other ALS inhibitor herbicides such as chlorsulfuron, imazamox and sulfometuron. Accession 03-1, however, did not show multiple resistance to the ACCase inhibitor herbicides diclofop, fluazifop, clethodim, sethoxydim and pinoxaden, nor to glyphosate. The in vivo ALS activity of the 03-1 biotype was less affected by mesosulfuron-methyl than the susceptible biotype. This indicates that the resistance mechanism of Italian ryegrass to mesosulfuron-methyl is partly due to an alteration in the target enzyme, ALS. It is concluded that diclofop-resistant ryegrass in the southern United States can be generally controlled by mesosulfuron-methyl. However, mesosulfuron-methyl must be used with caution because not all ryegrass populations are susceptible to it. There is a need for more thorough profiling of ryegrass resistance to herbicides.     

Propanil and swep inhibit 4-coumarate:CoA ligase activity in vitro

4-Coumarate:CoA ligase (4CL, EC 6.2.1.12) in the phenylpropanoid pathway in plants has attracted interest as a novel target for developing effective plant growth inhibitors (PGIs). In a previous study in which the 4CL inhibitory activity of 28 existing herbicides was investigated using an optimized in vitro screening assay, 4CL activity was found to be strongly inhibited by propanil and swep at 100 microM. Here, further experimental evidence is provided to substantiate the previous result. Using 4-coumaric acid as substrate, tobacco 4CL activity was inhibited by propanil or swep in a concentration-dependent manner, with 50% inhibition concentrations (I(50)) of 39.6 and 6 microM respectively. These herbicides also exhibited uncompetitive inhibition towards 4-coumaric acid. Furthermore, 4CLs from several plant species were inhibited by the herbicides within a range from 1 to 50 microM. It is proposed that these herbicides have another site of action as a result of the inhibition of 4CL in the phenylpropanoid pathway, and this enzyme represents a new target site for the development of PGI.     

氮素调控对玉米氮素同化过程及产量的影响

氮素调控措施与作物氮素吸收利用和产量密切相关,但目前关于不同氮素调控措施对玉米主要生育期氮素同化过程的影响仍不清楚。以郑单958为试验材料,设置不施氮肥（CK）、传统施肥（CN）、氮肥+生物炭（SN）和氮肥+硝化抑制剂DMPP（DN）4个处理,分析不同氮素调控对玉米氮素同化过程中铵态氮和硝态氮含量、硝酸还原酶（NR）和谷氨酰胺合成酶（GS）活性、游离氨基酸和可溶性蛋白含量以及氮素利用率和产量的影响。结果表明：DN和SN处理较传统施肥处理均可以提高玉米植株体内硝态氮和铵态氮含量、NR和GS活性;DN和SN处理显著提高灌浆期谷氨酸、游离氨基酸和可溶性蛋白含量;DN处理成熟期籽粒的氮素积累量显著高于SN和CN处理,分别显著增加18.4%和30.0%;DN处理产量最高,SN次之,二者并无显著差异,但相较CN处理分别显著增产1 483.0和1 154.2 kg·hm^-2。两种氮素调控均促进了玉米对氮素的吸收,显著提高氮肥吸收利用率,其中硝化抑制剂处理氮肥吸收利用率最高且显著高于其他处理。综上,生物炭或硝化抑制剂配施氮肥,可以促进玉米氮素同化和转运过程,显著提高玉米产量和氮肥利用效率,综合产量、籽粒氮素积累量和氮肥吸收利用率,硝化抑制剂配施氮肥可作为淮河流域玉米高产高效的栽培措施。