In this study we analysed the influence of drought stress on the leaf morphological characteristics, osmotic adjustment substances, antioxidant enzymes, and resistance-related photosynthetic physiological indices of Harumi tangor plants grafted onto Poncirus trifoliata (Pt), Citrus junos (Cj), and Citrus tangerine (Ct). The leaf relative water content and leaf area of the three rootstocks decreased with increasing drought stress, with the smallest decrease in Cj. The relative conductivity and malondialdehyde content increased with increasing drought stress. Proline, total soluble sugar, soluble protein, and activities of superoxide dismutase, ascorbate peroxidase, and catalase increased with drought stress but decreased under severe drought stress, with Cj exhibiting the greatest increase in enzyme activity. The net photosynthetic rate, stomatal conductance, transpiration rate, and chlorophyll a and b content were all lower than those of the control, whereas intercellular CO2 concentration increased with increasing drought stress. The initial fluorescence and maximal quantum yield of PSII were approximately equal for all rootstocks but increased with increasing drought stress severity. The combined analysis of physiological indicators, membership function, and principal components indicated that the drought resistance of grafted H. tangor decreased in the order Cj > Ct > Pt.
Considering the known effects of xyloglucan endotransglycosylase (XET) on plant growth and development, we aimed to determine whether XETs help to regulate the growth and elongation of Huangguogan shoots and roots. We confirmed a possible role for XET during seedling etiolation. Our results revealed that the roots of etiolated seedlings (H-E) were longer than those of green seedlings (H-G). However, shoot length exhibited the opposite pattern. We also observed positive and negative effects on the xyloglucan-degrading activity of XET in the root sub-apical region and shoots of etiolated Huangguogan seedling, respectively. There was a significant down-regulation in CitXET expression in the etiolated shoots at 15 days after seed germination. On the contrary, it was significantly increased in the root sub-apical region of etiolated and multicolored seedlings at 15 days after seed germination. The XET coding sequence (i.e., CitXET) was cloned from Huangguogan seedlings using gene-specific primers. The encoded amino acid sequence was predicted by using bioinformatics-based methods. The 990-bp CitXET gene was highly homologous to other XET genes. The CitXET protein was predicted to contain 319 amino acids, with a molecular mass of 37.45 kDa and an isoelectric point of 9.05. The predicted molecular formula was C1724H2548N448O466S14, and the resulting protein included only one transmembrane structure. The CitXET secondary structure consisted of four main structures (i.e., 21% α-helix, 30.72% extended strand, 9.09% β-turn, and 39.18% random coil). Analyses involving the NCBI Conserved Domains Database (NCBI-CDD), InterPro, and ScanProsite revealed that CitXET was a member of the glycosyl hydrolase family 16 (GH16), and included the DEIDFEFLG motif. Our results indicate that the differed degrees of etiolation influenced the CitXET expression pattern and XET activity in Huangguogan seedlings. The differential changes in XET activity and CitXET expression levels in Huangguogan seedlings may influence the regulation of root and shoot development, and may be important for seedling etiolation.
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