Protein degradation and fermentation characteristics of unwilted red clover and alfalfa silage harvested at various times during the day

Extensive proteolysis during fermentation of high‐protein legumes reduces dietary N‐use efficiency in ruminants. Research has demonstrated that enhancing the level of fermentable carbohydrates in crops entering the silo may reduce protein degradation by increasing the rate of decline in pH. The objective was to evaluate whether delaying cutting time during the day, to allow accumulation of total non‐structural carbohydrates (TNC), would inhibit proteolysis in the silo. Red clover (Trifolium pratense L.) and alfalfa (Medicago sativa L.) were harvested at 06.00, 10.00, 14.00 and 18.00 hours in 1993, 1994 and 1995, and ensiled without wilting. TNC accumulated in fresh forage during the day, with starch accounting for more than 0·50 of the daily change in TNC in fresh herbage of both species, except in red clover in 1995. The level of TNC in fresh forage did not consistently affect the extent of protein degradation in either species and, in all instances, alfalfa underwent more extensive proteolysis than red clover. Silage pH typically decreased and starch concentration increased as cutting time was delayed from 06.00 to 18.00 hours. Although the extent of proteolysis was largely unaffected by inherent increases in TNC, harvesting in the afternoon did provide several benefits including increased dry‐matter content, lower silage pH and higher starch concentrations. Effluent production is a concern in any unwilted silage system; there was therefore an added advantage of lower moisture content from cutting in the afternoon.     

Interaction between water-soluble carbohydrate reserves and defoliation severity on the regrowth of perennial ryegrass (Lolium perenne L.)-dominant swards

A field-study was undertaken in Hamilton, New Zealand to determine if there was an interaction between water-soluble carbohydrate (WSC) reserve content and defoliation severity on the regrowth of perennial ryegrass-dominant swards during winter. Perennial ryegrass plants with either low or high WSC content were obtained by varying the defoliation frequency. At the third defoliation at the one-leaf stage and at the first defoliation at the three-leaf stage (harvest H₁), swards were mown with a rotary lawnmower to residual stubble heights of 20, 40 or 60 mm. All swards were then allowed to regrow to the three-leaf stage before again defoliating to their treatment residual stubble heights (H₂). Frequently defoliated plants contained proportionately between 0·37 and 0·48 less WSC in the stubble after defoliation, depending on the severity of defoliation. There was no interaction between WSC content and defoliation severity for herbage regrowth between harvests H₁ and H₂. Herbage regrowth was lower from swards containing low WSC plants compared with high WSC plants (2279 vs. 2007 kg DM ha⁻¹). Furthermore, swards defoliated to 20 or 40 mm had greater herbage regrowth compared with those defoliated to 60 mm (2266, 2249 and 1914 kg DM ha⁻¹ for swards defoliated to residual stubble heights of 20, 40 and 60 mm, respectively). Regrowth of perennial ryegrass was positively correlated with post-defoliation stubble WSC content within defoliation severity treatment, implying that WSC contributed to the defoliation frequency-derived difference in herbage yield. However, the effect of defoliation severity on herbage regrowth was not associated with post-defoliation stubble WSC content.     

栓皮栎体胚的增殖、成熟和萌发

以栓皮栎实生苗叶片为外植体诱导体细胞胚胎发生,调查碳水化合物、渗透剂和植物生长调节剂对体胚增殖、成熟和萌发的影响,建立体胚增殖、成熟和萌发的培养基方案.叶片外植体在附加1.0 mg·L-1NAA和0.5mg·L-1BA的起始培养基上诱导形成前胚性团块.这些胚性团块在增殖培养基上培养6周,在附加1 mg·L-1BA、0.25 mg·L-1NAA和3%蔗糖的MS培养基上增殖效果最好.将单个体胚转接到成熟培养基上进行培养,蔗糖浓度对栓皮栎体胚成熟以及后续的萌发有显著影响.成熟培养基中附加5%的蔗糖,体胚成熟率和萌发率分别达到63.5%和33.8%.虽然在成熟培养基中附加ABA有利于体胚成熟,但对体胚的进一步萌发没有促进作用.为了提高萌发率,成熟体胚在附加植物生长调节剂的萌发培养基上进行培养,以及进行预冷处理.成熟体胚4℃冷处理没有促进胚根和上胚轴的发育萌发.在附加0.5 mg·L-1BA和0.25 mg·L-1 IBA的1/2 MS萌发培养基上,体胚萌发率达到65.9%,再生植株转化率达到9.4%.     

猪繁殖与呼吸综合征病毒非结构蛋白研究进展

猪繁殖与呼吸综合征(PRRS)是由猪繁殖与呼吸综合征病毒(PRRSV)引起的高度接触性传染病。主要引起母猪繁殖障碍,表现为妊娠后期流产,死胎,木乃伊胎及弱仔;各日龄段的猪特别是仔猪感染PRRSV引起呼吸道疾病,以间质性肺炎为特征,论文主要从近几年研究的PRRSV的非结构蛋白特性、免疫学功能、非结构蛋白基因的变异等方面进行分析,阐述非结构蛋白在病毒的增殖、致病力产生的影响,从而为进一步的研究提供一定的理论基础。     

禽呼肠病毒P10、P17非结构蛋白基因的克隆及序列分析

根据GenBank上的禽呼肠病毒(ARV)S1基因序列,设计并合成了一对跨越P10和P17非结构蛋白基因的特异性引物,对13个ARV毒株进行RT-PCR扩增、克隆及序列测定。结果显示,13个ARV毒株的P10蛋白基因ORF全长均为297bp,编码98个氨基酸;P17蛋白基因ORF全长为441bp,编码146个氨基酸。这13个ARV毒株P10、P17蛋白基因核苷酸同源性分别在96.6%~100%和95.2%~99.3%之间,推导的氨基酸同源性分别在98.2%~100%和91.9%~99.0%之间。将这13个ARV毒株与GenBank上其他正呼肠病毒毒株,包括番鸭株(DRV)和飞狐上分离到的内尔森海湾病毒(NelsonBayvirus,NBV)及两个澳洲分离株(ARM-1和SOM-4)进行同源性比较和遗传进化树分析,结果表明,呼肠病毒有地域和种类的差别。     

禽呼肠孤病毒P17蛋白基因在大肠杆菌中的表达及其ELISA检测方法的建立

用设计合成的1对跨越禽呼肠孤病毒（ARV）P17非结构蛋白基因完整开放阅读框（ORF）的特异性引物，对ARVS1133株进行了RT-PCR扩增。扩增产物与pGEX-4T-1原核表达载体连接后，转化大肠杆菌BL21感受态细胞。经0．2mmol／LIPTG诱导，表达的融合蛋白分子质量为42.4ku，占茵体总蛋白的34％。表达产物P17蛋白经不同浓度尿素纯化后，纯度达到85％。Western-blotting显示，纯化的P17蛋白能与感染ARV的阳性血清反应，说明其具有抗原性。以此重组蛋白为包被抗原初步建立了用于鉴别检测ARV活病毒感染与灭活疫苗免疫的SPF鸡血清的ELISA。     

Removal of flowers or inflorescences affects ‘Barnea’ olive fruitlet post-anthesis abscission

A typical olive (Olea europaea L.) inflorescence consists of about 20 flowers. However, in many cultivars, only one fruit develops. This is due to massive abscission of flowers and fruitlets, which occurs during the first month after anthesis. In this study, we used the olive cultivar 'Barnea' to characterize the abscission mechanism and to try to increase fruit set by increasing the number of developed fruit per inflorescence. Removing the lateral flowers 3 weeks before anthesis increased fruit set by more than 50%. Removing all inflorescences but one from a branch increased the number of developed fruits from 0.93 to 2.8 during 2017 and from 0.91 to 3.34 fruits per inflorescence in 2018. Sugar quantification in the pistil revealed that starch level is high on the day of anthesis and low 25 days later in abscised as well as in developed fruit. Soluble carbohydrates are low on the day of anthesis, low in abscised flowers/fruitlets 25 days after anthesis and high in developed fruit. Screening the natural variation found in the Israeli germplasm collection revealed that in most cultivars less than one fruit per inflorescence has developed. However, there are unique cultivars with a higher fruit set.     

水稻灌浆结实期籽粒接受14C-同化产物能力的研究

在盆栽条件下，以粳稻日本晴为材料，于抽穗后5、20、35d整株标记^14CO2 1．5h，同化后24h取样，测定根、叶片、茎鞘、穗轴、枝梗、小枝梗及籽粒中^14C-同化产物的淀粉量和可溶性糖含量。结果表明：随着抽穗后天数的增加，全株^14C-同化量明显减少，光合强度下降的幅度大于绿叶重；灌浆初期固定的^14C-光合产物在同化后24h60％以上被运输到籽粒中，籽粒中^14C-光合产物的淀粉量和可溶性糖含量均明显高于其他器官，证明灌浆初期籽粒具有主动接受^14C-光合产物的能力；灌浆后期叶片、茎鞘、穗轴、枝梗、小枝梗中含有大量的^14C-同化产物，且多为能运输的可溶性糖，但籽粒中^14C-光合产物很少，说明灌浆后期籽粒接受光合产物能力明显变弱，是造成大量^14C-光合产物滞留在营养器官中的重要原因；籽粒的含水率和淀粉合成能力与籽粒接受^14C-光合产物量之间具有明显的伴随关系，推测籽粒的含水率和淀粉合成能力低可能是造成籽粒接受光合产物能力下降的重要原因之一。     

猪繁殖与呼吸综合征病毒Nsp2蛋白纳米抗体的筛选及其鉴定

为制备猪繁殖与呼吸综合征病毒(PRRSV)非结构蛋白2(Nsp2)的纳米抗体,本研究将截短表达的Nsp2重组蛋白免疫骆驼后分离骆驼外周血淋巴细胞,利用RT-PCR扩增其VHH基因,将VHH基因插入pCANTAB5E噬菌体载体,构建双峰驼重链抗体可变区文库。利用噬菌体展示技术经过3轮淘选,共筛选得到44株Nsp2特异性纳米抗体;经ELISA检测,结果显示44株Nsp2特异性纳米抗体均能与Nsp2蛋白特异性反应。本研究首次筛选到PRRSVNsp2特异性纳米抗体,为Nsp2蛋白的相关研究提供了必要的研究工具,同时为开发新型抗PRRSV药物奠定一定的基础。