Traditional breeding processes to genetically modify the long reproductive cycle and slow seed maturation of orchids have limits. We developed a more efficient protocol using particle bombardment to produce transgenic plants of Oncidium Sharry Baby 'OM8' (Orchidaceae). Pretreating protocorm-like bodies (PLBs) with 0.5 M sucrose for 2 h increased single-cell embryogenesis 3-to 4-fold; however, shoot formation was suppressed. In addition, new PLBs were regenerated from the entire sucrose-pretreated PLBs, whereas in untreated PLBs, this occurred only from the bases. Pretreated PLBs were bombarded with pSPFLP containing genes encoding a sweet pepper ferredoxin-like protein (pflp), hygromycin phosphotransferase (hpt) and b-glucuronidase (GUS) driven by the cauliflower mosaic virus 35S promoter. Pretreated PLBs showed a 14.8-fold increase in GUS expression over the untreated PLBs 40 days after bombardment. The presence of pflp and hpt transgenes in the 40 putatively stably transformed lines that produced 113 clones was confirmed by PCR analysis. Six lines (eight clones) were positive for both pflp and hpt transgenes. In addition, clones derived from these lines were either all positive or all negative for the two transgenes, which suggests homogeneity in pretreated PLBs with more single-cell embryogenesis. Thus, sucrose pretreatment enhanced the regeneration of PLBs, single-cell embryogenesis and efficiency of transformation.Abbreviations: 35S promoter -cauliflower mosaic virus 35S promoter; GUS -b-glucuronidase; hpthygromycin phosphotransferase gene; nos terminator -terminator of nopaline synthase gene; pflp -pepper ferredoxin-like protein sequence; PLB -protocorm-like body; uidA -b-glucuronidase gene

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The relationships between carbon isotope discrimination (Δ) and some morphophysiological traits such as specific leaf dry weight (SLDW), gas exchange parameters, and relative water content (RWC) were studied in a collection of 20 bread wheat cultivars (landraces, released cultivars and advanced lines) in three locations of the Ningxia region (North-East China), i.e. Yinchuan (limited irrigation conditions), Huinong (limited irrigation conditions + salinity) and Guyuan (rain-fed conditions). Relationships between Δ, grain yield (GY), and harvest index (HI) and above-ground biomass (AGB) were also analysed. Differences in the measured traits between different locations were highly related to the variation in water availability. Positive correlations were noted between Δ and HI and grain yield. Flag leaf Δ was positively correlated with RWC at anthesis, and negatively associated with SLDW at grain filling. Significant and negative correlations between Δ and dry matter weight per plant at anthesis and biomass at maturity were noted. Leaf temperature (LT) was found to be negatively correlated with Δ and gs. The findings suggest that Δ may be a useful indicator reflecting wheat yield, harvest index, and water status under irrigation and rain-fed conditions in the Ningxia region.

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