Sword-leaf dogbane (Apocynum venetum) is a traditional Chinese herb with increasingly recognized potential to enhance health, but no study of stable reference genes in this herb has been reported. Based on a homologous cloning strategy, we have successfully cloned five candidate reference genes from sword-leaf dogbane: glyceraldehyde-3-phosphate dehydrogenase (AvGAPDH), beta tubulin (AvbTUB), polyubiquitin (AvUBQ), elongation factor 1-alpha (AvEF1α), and actin (AvACTIN). Three distinct algorithms, geNorm, NormFinder, and BestKeeper, were used to estimate the expression stability of candidate reference primer pairs. We found that AvACTIN-2 and AvACTIN-3 presented the highest stability of expression in different tissue samples, and AvGAPDH-2 was most stable under salinity stress. In addition, we illustrated the application of these new reference genes by assaying the expression levels of two hyperoside biosynthesis terminal enzyme genes, flavonoid 3′-hydroxylase (F3′H) and flavonol synthase (FLS), under salinity stress. Our study is the first to report stable expression of internal reference genes in sword-leaf dogbane in multiple experimental sample sets.
Zongchang Xu, Meng Wang, Jinhui Zhou, Han Liu, Chengsheng Zhang, and Yiqiang Li
Wanmei Jin, Jing Dong, Yuanlei Hu, Zhongping Lin, Xuefeng Xu, and Zhenhai Han
Dehydration response element binding (DREB)1b is a cold-inducible transcription factor in Arabidopsis thaliana. DREB1b driven by cauliflower mosaic virus 35S promoter was genetically introduced into grape Vitis vinifera L. cv. Centennial Seedless through Agrobacterium-mediated transformation for improving its cold resistance and exploring new genetic breeding approaches to obtain cold-resistant cultivars. In this study, Southern blot analysis showed the DREB1b gene was integrated into the transgenic grapevines with one to two copies. Northern blot analysis showed the presence of DREB1b transcripts in the independent transgenic lines 3, 5, 6, and 7. Further characterization of transgenic grapevines confirmed that both electrolyte leakage conductivity and the freezing point of the transgenic plants were lower than those of wild-type plants. After the cold treatment at –4 °C for 12 h, 26% of transgenic plants wilted among which 95% plants recovered once being placed under the condition of temperature 22 to 25 °C. However, subjected to the same treatment, 98% of nontransgenic plants wilted and only 2% recovered. Our results lead to the conclusion that activity of DREB1b in the transgenic grape could significantly improve its resistance to cold stress.
Chenping Zhou, Ruiting Chen, Yaqiang Sun, He Wang, Yi Wang, Ting Wu, Xinzhong Zhang, Xuefeng Xu, and Zhenhai Han
Bridge grafting is widely applied in trunk-wounded apple trees. In this study, we carried out semigirdling and ring girdling on the trunk of ‘Nagafu 2’/Malus baccata (L.) Borkh apple trees to simulate trunk injury. We then bridge grafted a M9 self-rooted rootstock on the injured trunks to study the effects of bridge grafting on flowering, fruit-set, tree vigor, and fruit characteristics in ‘Nagafu 2’ apple. The results showed that both semigirdling and ring girdling due to the large wounded area caused significant decrease in flowering, fruit-set, and tree vigor (estimated by measuring leaf area, leaf gas exchange, tree height, and shoot growth); in addition, ring girdling increased flesh and peel firmness. However, bridge grafting of M9 self-rooted rootstock on semigirdling and girdling apple trees resulted in partial recovery of tree vigor (leaf area and photosynthesis) and maintaining the reduction of vegetative growth, thereby increasing flowering, fruit-set, yield, fruit weight, and peel firmness.