Improving the poor resistance to environmental stress and the weak development of roots system in the cherry (Prunus) rootstock ‘Gisela 6’ (Prunus cerasus × Prunus canescens) is of great importance for sustainable sweet cherry (Prunus avium) production. Although a stable genetic transformation system has been developed for ‘Gisela 6’ rootstock, there is little information on the identification of genes involved in stress resistance. Using the cherry rootstock cultivar Gisela 6, we identified a total of 12 novel mitogen-activated protein kinase (MAPK) genes, designated PcMPKs. Phylogenetic analysis revealed that the PcMPKs could be divided into four groups, designated A, B, C, and D. In addition, an intron–exon structure analysis for the PcMPKs was conducted to help further understand the structure–function relationships within the cherry family. The expression profiles of PcMPKs in response to abiotic and biotic stresses were characterized using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Five PcMPKs (i.e., PcMPK4-1, PcMPK4-2, PcMPK3, PcMPK6, and PcMPK18) exhibited differential expression, and suggested their potential roles in plant responding to various stresses. This study provides the basis for further analysis on the physiological functions of PcMPKs in environmental tolerance in cherry rootstocks.
Xiaojuan Zong, Jiawei Wang, Li Xu, Hairong Wei, Xin Chen, Dongzi Zhu, Yue Tan and Qingzhong Liu
Xiaojuan Zong, Brandon J. Denler, Gharbia H. Danial, Yongjian Chang and Guo-qing Song
‘Hansen 536’ (Prunus dulcis × Prunus persica) is an important commercial rootstock for peach and almond. However, susceptibility to wet soil and bacterial canker has limited its use primarily to areas with less annual rainfall. Genetic engineering techniques offer an attractive approach to improve effectively the current problems with this cultivar. To develop an efficient shoot regeneration system from leaf explants, 10 culture media containing Murashige and Skoog (MS) or woody plant medium (WPM) supplemented with different plant growth regulators were evaluated, and adventitious shoot regeneration occurred at frequencies ranging from 0% to 36.1%. Optimal regeneration with a frequency of 32.3% to 36.1% occurred with WPM medium containing 8.88 µm 6-benzylamino-purine (BAP) and 0.98 to 3.94 µm indole-3-butyric acid (IBA). The regenerated shoots had a high rooting ability, and 80% of the in vitro shoots tested rooted and survived after being transplanted to substrate directly. Transient transformation showed an efficient delivery of the β-glucuronidase (GUS) reporter gene (gusA) using all three Agrobacterium tumefaciens strains tested with a concentration of OD600 0.5 to 1.0 for 4 days of cocultivation. The protocols described provide a foundation for further studies to improve shoot regeneration and stable transformation of the important peach and almond rootstock ‘Hansen 536’.