In order to provide theoretical references for early-maturing Citrus unshiu Marc. in terms of fruit thinning, timely fertilization and harvesting, as wel as for for-mulation of of Early-maturing Citrus unshiu Marc. Cu...In order to provide theoretical references for early-maturing Citrus unshiu Marc. in terms of fruit thinning, timely fertilization and harvesting, as wel as for for-mulation of of Early-maturing Citrus unshiu Marc. Cultivation Technique Standards in Guizhou Province, the dynamic changes of fruit longitudinal diameter and trans-verse diameter of 11-year-old and 12-year-old early maturing Citrus unshiu Marc. were tested in two consecutive years and the effects of single-fruit bearing branch on fruit growth and development were compared. The results showed annual varia-tion of longitudinal and transverse diameter of the fruit in Guizhou grew faster at the beginning and then slowed down gradual y, maintaining a slow-growth period after October; the growth curve final y was flat in mature stage. General y, the growth and development stage of fruit showed four growth peaks, showing an in-creasing S-shape curve and fruit net increase also had four growth peaks with a double S-shape curve. Specifical y, longitudinal diameters growth alternated with transverse diameter growth and the former proved much stronger. In terms of ef-fects of single-fruit bearing branch on longitudinal and transverse diameter growth, the number of reserved fruits had direct effects on the growth, net increase of weight, time of growth peak, fluctuation and the number of growth peak. The growth and development, and quality of fruits in the treatments with two or three fruits re-served on single fruit-bearing branch proved better compared with the control with four left fruits. It suggested that early June and early July are the best time for fer-tilization of early-maturing Citrus unshiu Marc. in Guizhou Pprovince and would have better effect if combined with foliage spraying. The suitable harvest time is in the end of October. The preferred number of reserved fruit on single fruit-bearing branch is 2-3. In actual production, the implementation of flower and fruit thinning should also takes tree age, balance of tree vigor, strength of fruit bearing branch, alternate bearing phenomenon and other factors into consideration.展开更多
为了给深入研究猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndromevirus,PRRSV)ORF6基因编码的M蛋白的生物学功能提供重要试验材料,本研究首先利用慢病毒包装系统构建了过表达PRRSVORF6基因的重组慢病毒质粒,将该...为了给深入研究猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndromevirus,PRRSV)ORF6基因编码的M蛋白的生物学功能提供重要试验材料,本研究首先利用慢病毒包装系统构建了过表达PRRSVORF6基因的重组慢病毒质粒,将该质粒连同辅助质粒共同转染至HEK293T细胞获得重组慢病毒;之后将重组慢病毒感染MARC-145细胞,利用嘌呤霉素结合有限稀释法进行筛选,连续筛选3轮后建立了稳定表达PRRSVM蛋白的MARC-145ORF6细胞系;并使用CCK-8试验评估过表达PRRSVM蛋白对MARC-145细胞生长的影响。利用RT-PCR、蛋白免疫印迹(Westernblot)和间接免疫荧光(IFA)评估MARC-145ORF6细胞系的传代稳定性并鉴定M蛋白的亚细胞定位,进一步利用RT-qPCR评估过表达M蛋白对MARC-145细胞的干扰素及相关调节基因的影响;此外,还测定了PRRSV在MARC-145ORF6细胞系、MARC-145Flag细胞系和MARC-145细胞中的病毒滴度并绘制多步生长曲线以比较其差异。CCK-8试验结果表明,过表达PRRSVM蛋白对MARC-145细胞活力无显著影响;RT-qPCR、Westernblot和IFA等试验结果表明,MARC-145ORF6细胞系能够表达PRRSV的M蛋白且在传代过程中稳定。此外,稳定表达PRRSVM蛋白显著下调了细胞系的Ⅰ型干扰素及其相关调节基因;多步生长曲线表明,MARC-145ORF6细胞系促进PRRSV增殖,提高其病毒滴度。综上,本研究构建了可以稳定表达PRRSVM蛋白的MARC-145ORF6细胞系,发现其Ⅰ型干扰素水平显著下调且促进PRRSV复制。本研究构建的MARC-145ORF6细胞系将为M蛋白功能的深入研究提供重要生物材料。展开更多
文摘In order to provide theoretical references for early-maturing Citrus unshiu Marc. in terms of fruit thinning, timely fertilization and harvesting, as wel as for for-mulation of of Early-maturing Citrus unshiu Marc. Cultivation Technique Standards in Guizhou Province, the dynamic changes of fruit longitudinal diameter and trans-verse diameter of 11-year-old and 12-year-old early maturing Citrus unshiu Marc. were tested in two consecutive years and the effects of single-fruit bearing branch on fruit growth and development were compared. The results showed annual varia-tion of longitudinal and transverse diameter of the fruit in Guizhou grew faster at the beginning and then slowed down gradual y, maintaining a slow-growth period after October; the growth curve final y was flat in mature stage. General y, the growth and development stage of fruit showed four growth peaks, showing an in-creasing S-shape curve and fruit net increase also had four growth peaks with a double S-shape curve. Specifical y, longitudinal diameters growth alternated with transverse diameter growth and the former proved much stronger. In terms of ef-fects of single-fruit bearing branch on longitudinal and transverse diameter growth, the number of reserved fruits had direct effects on the growth, net increase of weight, time of growth peak, fluctuation and the number of growth peak. The growth and development, and quality of fruits in the treatments with two or three fruits re-served on single fruit-bearing branch proved better compared with the control with four left fruits. It suggested that early June and early July are the best time for fer-tilization of early-maturing Citrus unshiu Marc. in Guizhou Pprovince and would have better effect if combined with foliage spraying. The suitable harvest time is in the end of October. The preferred number of reserved fruit on single fruit-bearing branch is 2-3. In actual production, the implementation of flower and fruit thinning should also takes tree age, balance of tree vigor, strength of fruit bearing branch, alternate bearing phenomenon and other factors into consideration.
文摘为了给深入研究猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndromevirus,PRRSV)ORF6基因编码的M蛋白的生物学功能提供重要试验材料,本研究首先利用慢病毒包装系统构建了过表达PRRSVORF6基因的重组慢病毒质粒,将该质粒连同辅助质粒共同转染至HEK293T细胞获得重组慢病毒;之后将重组慢病毒感染MARC-145细胞,利用嘌呤霉素结合有限稀释法进行筛选,连续筛选3轮后建立了稳定表达PRRSVM蛋白的MARC-145ORF6细胞系;并使用CCK-8试验评估过表达PRRSVM蛋白对MARC-145细胞生长的影响。利用RT-PCR、蛋白免疫印迹(Westernblot)和间接免疫荧光(IFA)评估MARC-145ORF6细胞系的传代稳定性并鉴定M蛋白的亚细胞定位,进一步利用RT-qPCR评估过表达M蛋白对MARC-145细胞的干扰素及相关调节基因的影响;此外,还测定了PRRSV在MARC-145ORF6细胞系、MARC-145Flag细胞系和MARC-145细胞中的病毒滴度并绘制多步生长曲线以比较其差异。CCK-8试验结果表明,过表达PRRSVM蛋白对MARC-145细胞活力无显著影响;RT-qPCR、Westernblot和IFA等试验结果表明,MARC-145ORF6细胞系能够表达PRRSV的M蛋白且在传代过程中稳定。此外,稳定表达PRRSVM蛋白显著下调了细胞系的Ⅰ型干扰素及其相关调节基因;多步生长曲线表明,MARC-145ORF6细胞系促进PRRSV增殖,提高其病毒滴度。综上,本研究构建了可以稳定表达PRRSVM蛋白的MARC-145ORF6细胞系,发现其Ⅰ型干扰素水平显著下调且促进PRRSV复制。本研究构建的MARC-145ORF6细胞系将为M蛋白功能的深入研究提供重要生物材料。
