Malus hupenensis var. `Pingyitiancha' is an important apple stock with many good characteristics, including waterloggig resistance, cold resistance, salt resistance and so on. The three group gene-HVA1 come from barley was transformed into `Pingyitiancha' mediated by Agrobacterium tumefaciens and transformed regeneration plants were obtained in this research. The HAV1 gene cloned from plasmid containing it (offered by Dr. Guo Weidong) by PCR with high fidelity pfu Taq DNA polymerase. It was ligated between BamH 1 and Sac 1 site in PUC118 vector, and identified by electrophoresis after digested with BamH 1 and Sac 1. Through nuclear sequence detecting, it is confirmed that the HAV1 gene cloned in this research is 703bp.This fragment was ligated with 11kb fragment from pB121 plasmid and constructed pBHA vetor. The pBHA vector was introduced in A.tum LBA4404 by triparental mating and the binary vector was obtained. It is cinfirmed that HVA1 gene had been insert in T-DNA by in situ hybirdization. Using `Pingyitiancha' shoot apex, mediated by A. tum. System, the HAV 1 gene was transformed into the plant. Kam resistance regeneration plants were obtained, 6 of them were confirmed as transformation plants by PCR and dot blot.
Xian Shen*, Ling Guo and Zhenlin Wei
Xiang Shen*, Zhenlin Wei and Ling Guo
`Pingyitiancha' has good agricultural traits and is an important rootstock for apples. It was studied for Kam, Cef and Carb concentrations, precultural times, active medium of A.tum, Ca++ concentration, mixed inoculation times of AgNO3, and PVP delay selection times for getting transgenic `Pingyitiancha' stock with more resistance to drought and salt. It was demonstrated that the highest transformation frequency was obtained with 200 mg·L-1 Cef as an A.tum restraining antibiotic, 8 mg·L-1 Kam as a selection antibiotic, GCJ8 active medium, a precultural of 2 days, 5 Ca++ concentrations in preculture and coculture medium compared with standard MS medium, immersed in A. tum as OD600 for 0.5 to 1 min., 4 mg·L-1 AgNO3 and 0.7% PVP as an anti-oxidation compound to reduce hydroxybenzene oxidation and delay selection for 5 days by using `Pingyitiancha' apex shoots as explants. Using mediate of A. tum strain LBA4404 under conditions mentioned above and same explants, the HVA1 gene were transformed into `Pingyitiancha' under the control of CaMV35S promoter and obtained Kam resistance regeneration plants. Six transgenic plants were confirmed by PCR and subsequent dot blot methods.
Wei-Ling Guo, Yao-Chien Alex Chang and Chien-Yuan Kao
Cyrtopodium paranaense is a tropical terrestrial orchid, which propagates mainly through sexual seed germination. In this study, we document the asexual morphogenesis of the root tip to protocorm-like body (PLB) conversion in Cyrtopodium paranaense. Protocorm-like bodies sporadically developed from root tips of flask-grown seedlings in the absence of any exogenous plant growth regulators (PGRs). The compact PLBs ultimately gave rise to normal plantlets. Histological observations revealed that the root cap became dissociated from the root apex at an early stage followed by dispersed extension of root vascular strands into nascent PLBs. Protocorm-like bodies also developed from the root central stele tissue. In root tip segment cultures, PLBs were not formed without providing PGRs but were efficiently formed from root tips in Murashige and Skoog (MS) medium supplemented with 10.2 μM indole-3-acetic acid (IAA) and 9.0 μM thidiazuron (TDZ). Both IAA and TDZ promoted the formation of PLBs; however, TDZ did not induce PLB formation in the absence of IAA, indicating a synergistic effect of the two PGRs. Protocorm-like bodies were proliferated and subsequently plants regenerated in PGR-free MS medium. Root tip culture may be used as an alternative method for the propagation of Cyrtopodium paranaense.
Haiying Zhang, Guoyi Gong, Shaogui Guo, Yi Ren, Yong Xu and Kai-Shu Ling
Because of the growing threat of global warming, drought stress could severely affect the normal growth and development of crop plants. To alleviate such an adverse effect, there is a need to screen watermelon germplasm collections to identify genetic sources for potential drought tolerance. In the present study, 820 accessions of USDA's Citrullus PIs and 246 watermelon breeding lines were evaluated for their drought tolerance at the seedling stage under extreme water stress conditions in a greenhouse. Significant variations in drought tolerance were observed in the Citrullus germplasm collections. Using fast clustering analysis, the tested watermelon materials could be assigned into four groups, including tolerant, intermediate tolerant, moderately sensitive, and sensitive, respectively. The most drought-tolerant Citrullus germplasm, including 13 Citrullus lanatus var. lanatus and 12 C. lanatus var. citroides accessions, were originated from Africa. These genetic materials could be used for rootstock breeding or for developing drought-tolerant watermelon cultivars.
Zhuping Fan, Yike Gao, Ling Guo, Ying Cao, Rong Liu and Qixiang Zhang
Bearded iris (Iris ×hybrida Hort.) is a large horticultural hybrid complex in the Iris genus, and the lack of understanding about its inheritance laws has seriously hindered the breeding process. From parental bearded iris ‘Indian Chief’ and ‘Sugar Blues’, four hybrid populations—including F1, F2, BC1P1, and BC1P2—were generated through hybridization. Fifteen key phenotypic traits, including plant height (PH), scape height (SH), length of fall (LF), width of fall (WF), length of standard (LS), width of standard (WS), and so on, were measured, and several genetic parameters (e.g., trait variation, heritability, trait correlation, distribution of flower color) were analyzed. The variation of phenotypic traits indicated that the F1 generation could produce larger flowers and a greater number of blooming stems than other generations, whereas backcrossing was beneficial at producing more flowers on one scape in the offspring of ‘Indian Chief’ and ‘Sugar Blues’. WF had the greatest broad-sense heritability (73.91%) among the 15 phenotypic traits, whereas the broad-sense heritability of SH was the lowest (2.06%). The correlation between a vegetative trait (PH) and a reproductive trait (WS) provided a path to early selection of germplasm. Furthermore, four important floral traits (LF, WF, LS, and WS) also correlated significantly to each other, thus simplifying the selection of larger flowers. Genes regulating fuchsia flower color were dominant over those for bluish purple flowers. Genetic effects of flower color in recurrent parents could be reinforced by backcrossing, thereby providing a potential way to modify flower color through hybridization.