The polymerase chain reaction (PCR) and RAPD fragments are potentially useful methods for identifying turfgrass cultivar breeding lines. RAPD markers were studied in 25 vegetatively propagated buffalograss lines using oligonucleotide random primers and agarose-gel electrophoresis to determine their potential for identifying cultivar breeding lines. The variation of RAPD markers was extensive. The RAPD markers produced by one random primer were sufficient to separate the 25 buffalograss lines. Cluster analysis baaed on' the RAPD markers produced by two random primers revealed that the 25 buffalograss lines generally fell into two groups: diploid and hexaploid. Three DNA extraction methods—sarcosyl lysis-chloroform extraction-isopropanol precipitation, sodium dodecyl sulfate (SDS) lysine-isopropanol precipitation, and boiling in the presence of Chelex-100 resin—and fresh or oven-dried tissues were tested for reproducibility of RAPD markers. The three DNA extraction methods, using dry or fresh plant tissues, produced highly comparable RAPD marker profiles. More than 80%1 of the RAPD markers was consistently detected in six replicate analyses. The above studies demonstrate that small quantities (5 mg) of oven-dried leaf tissue and several DNA extraction methods can be used for buffalograss fingerprint studies.
Echinacea purpurea L. is one of the important ornamental and medicinal plant species. Ploidy manipulation is a valuable tool for improving plant quality or production in E. purpurea as well as in many other plants. To study the segregation of pure ploidy plantlets from colchicine-induced ploidy chimeras in E. purpurea, we used a chimera plantlet that consisted of 1.93% diploid, 35.04% tetraploid, and 63.03% octoploid cells as the source material for experiments. The results showed that three factors significantly influenced the segregation, i.e., the component ratios of different ploidy cells in the chimera, the number of sequential passages, and the methods of segregation culture of the chimera plantlets. Other factors, such as explant types (i.e., leaf, petiole, or root) and 6-benzyladenine (BA) concentrations (i.e., 0.2, 0.4, 0.8, and 1.2 mg·L−1) occasionally influenced the segregation. Pure chromosome-doubled polyploids are not easily obtained in various plant species, so segregation culture of ploidy chimeras may potentially be more effective. The morphological characteristic and content of cichoric acid were compared among diploid, tetraploid, and octoploid plants. Results indicated that tetraploid and octoploid plants had more stunted growth, larger stomata, lower stomata frequency, more chloroplast number in guard cells, and higher cichoric acid content than original diploid lines.
Natural swimming pools (NSPs) rely on the interaction of bog vegetation, bacteria, and substrate to maintain water quality. Nitrogen (N) and phosphorus (P) levels in NSPs are critical because of their involvement in eutrophication. We conducted a 15-week greenhouse study to address the significant literature gap regarding nutrient removal capabilities of substrates and vegetation in the low-nutrient environment of NSPs. We used mass balance analyses to compare the performances of four substrates [river gravel (control), recycled glass, expanded clay, expanded shale] and two plant species [blue flag iris (Iris versicolor) and lizard’s tail (Saururus cernuus)] under two flow conditions: free water surface and subsurface flow. At the end of the experiment, except for the recycled glass group, all other substrate groups reduced water nitrate (NO3) levels to less than 30 mg⋅L−1, the standard of the 2011 Forschungsgesellschaft Landschaftsentwicklung Landschaftsbau (FLL) guidelines. However, only the expanded clay group closely approached the P standard (≤0.01 mg⋅L−1). Expanded clay and expanded shale demonstrated potential as substrates for NSP bogs. The final aboveground biomass dry weight was strongly negatively correlated with the final NO3 and P water concentrations. However, direct plant uptake proved insufficient to remove all nutrient inputs, especially for P. Except for the recycled glass group (34%), a significant portion of N (79%–90%) from total N added was removed by aboveground biomass. However, P uptake by biomass was substantially lower (18%–37%). With acceptable vigor and biomass accumulation, blue flag iris may be a suitable species for vegetated NSPs, whereas lizard’s tail is not because of uncertain establishment. Compared with controlling N, managing P for FLL standards in NSPs will be more challenging. Our work begins to fill the essential gap in the NSP literature regarding nutrient removal capabilities of substrates and vegetation. Future work should continue to study alternative substrates and plant species for P removal, particularly in field conditions and over longer periods.
The effects of high concentrations of Cl-, K+, Mg2+, and Ca2+ of the simulated waste water on the growth of turfgrass species and partitioning of these mineral element concentrations in the turfgrass-soil system have been studied. This is a two year project and the waste treatment was started in the first week of October 1993. The waste water contains 17.89 mM of K+, 97.5 mM of Ca2+, 78.1 mM of Mg2+, and 389.17 mM of Cl-. Kentucky bluegrass, perennial ryegrass, tall fescue, bermudagrass, and zoysiagrass have been irrigated with 1/5, 1/10, and 1/20 times concentration of the waste water and mowed weekly at 5 cm high. The preliminary results showed that there was no detectable growth inhibition of turfgrass by the three waste water concentrations. Waste water irrigation significantly increased the uptake of the mineral elements by the turfgrass. Significant reduction of the mineral element concentrations in the leach by the turfgrass system only found under the conditions of low concentration waste irrigation. However, the seasonal growth pattern of the turfgrass species may have significant influence on the partitioning of the element concentrations in the turfgrass-soil system and their concentrations in the leach. This prediction will be detected by the future studies.
The effects of regenerant wastewater irrigation and high concentrations of Ca2+, K+, Mg2+, and Cl– on growth and ion uptake of nine species of landscape plants were studied. Significant differences in chloride tolerance were detected among the species. Generally, the species that had greater uptake of chloride grew less than species that took up less amounts of chloride. Lace fern (Athyrium filix-femina Roth.) had the highest tissue Cl concentration and was the most affected. Hydrangea (Hydrangea macrophylla Ser.) also had high tissue Cl concentration, but showed no growth reduction. Its tolerance was attributable to a high tissue Ca concentration. The data suggest that in the species tested, higher tissue Ca concentrations were positively correlated with plant tolerance to Cl. Overall, the Cl– concentration in the wastewater seems to be the factor most likely to create problems for the landscape plants. However, severe negative effects will probably be noticed only for very sensitive plant species, but it is important to determine this before applying regenerant irrigation water.
Sorbitol is the primary photosynthate and translocated carbohydrate in apple (Malus ×domestica), and most of it is converted to fructose by sorbitol dehydrogenase (SDH) in sink tissues. We studied the expression of nine SDH genes, SDH activity, and sorbitol content of apple 1) in buds and floral tissues from dormancy to bloom, 2) in leaves and shoot tips of trees on two rootstocks, the moderately vigorous ‘Malling Merton 111’ (MM.111) and the dwarfing ‘Malling 9’ (M.9), and 3) in shoot tips in response to application of prohexadione-Ca to suppress shoot growth and defoliation and girdling (D/G) to deprive the shoot tip of sorbitol. In mature, orchard-grown trees, sorbitol was the main soluble carbohydrate in expressed xylem sap from dormancy to bloom at levels over 3- to 6-fold those of glucose and fructose, the other major sugars present. Sorbitol levels there increased from dormancy to its highest concentration at the half inch green stage and declined by bloom, while those of the other sugars increased. SDH activity per milligram of protein increased over 4-fold from dormancy to flowering. Three of the nine known SDH genes (SDH1, SDH2, and SDH3) were expressed in immature and mature leaves and all buds from dormancy to bloom, as well as in all floral organs, except that only SDH3 transcript was found in stamen tissue. Two genes, SDH6 and SDH9, were floral-tissue specific; SDH6 transcript was detected in all floral organs except stamens at full bloom, and SDH9 was only expressed in anthers with pollen. In buds and leaves of young, container-grown trees, SDH1 and SDH2 generally accounted for the majority of total SDH expression. There were generally no effects of rootstock on SDH expression, SDH activity, or sorbitol concentration in leaves, while apical shoot tips on M.9 rootstock exhibited greater SDH activity than those on MM.111 or lateral shoot tips on either rootstock, though SDH expression of apical and lateral shoot tips on M.9 was lower than on MM.111. Prohexadione-Ca reduced apical but not lateral shoot growth, increased apical but not lateral shoot tip sorbitol content, had no effect on SDH activity, and increased SDH1 expression of all shoot tips. D/G treatment reduced shoot growth, sorbitol content, and SDH activity, but increased SDH1 expression of apical shoot tips only and SDH2 expression of lateral shoot tips only. This work indicates that sorbitol and other sugars are abundantly available from dormancy to bloom, that SDH activity increased during this period, and that SDH expression is at least in part developmentally regulated within the individual floral and leaf tissues. In shoot tips and leaves of young trees, SDH transcript level was not correlated with sorbitol availability or SDH activity, suggesting that other factors have significant regulatory effects after SDH expression on SDH activity.
The most obvious effects of a low leaf:fruit (LF) ratio [two leaves for one cluster per shoot (LF2)] on grape (Vitis vinifera) berries are suppressed anthocyanin biosynthesis in the berry skin, decreased berry weight and soluble solids concentration, and increased titratable acidity. In this study, proteins isolated from berry skins grown under low and high LF ratio conditions, LF2 and LF12, respectively, were characterized by two-dimensional gel electrophoresis coupled to mass spectrometry. A survey of ≈600 to 700 spots from berry skin yielded 77 proteins with differential expression between LF12 and LF2 treatments. Of these, the 59 proteins that were identified consisted of 47 proteins that were down-regulated and 12 that were up-regulated under LF2 conditions compared with LF12 conditions. Most proteins involved in metabolism, energy, transcription, protein synthesis, binding function, signal transduction, and cell defense were down-regulated in LF2 berries, whereas two important enzymes of anthocyanin biosynthesis, chalcone synthase and dihydroflavonol reductase, were not detected. Only a few proteins (e.g., two heat shock proteins related to protein fate and nutrient reservoir storage protein) were found to be up-regulated in LF2 berries. This suggested that, with the exception of secondary metabolism, many proteomic events may have an effect on anthocyanin synthesis in the skins responding to LF.
Five peach cultivars [Prunus persica (L.) Batch] with different maturity dates were subjected to sink–source manipulation by girdling to isolate 1-year-old fruit-bearing shoots. Four treatments were performed: fruit were removed (−fruit); one fruit (+1 fruit) and two fruit (+2 fruit) were kept inside two girdling cuts; and two fruit were kept outside two girdling cuts (−fruit*). Photosynthetic responses for the five cultivars were similar and did not show genotypic differences. Generally, net photosynthetic rate (Pn), stomatal conductance (gs), and transpiration rate (E) were higher, and leaf temperature (Tl) was lower in +2 fruit than in +1 fruit, followed by −fruit and −fruit* which were not different. The results also indicated that water outflow from fruit into leaves did not influence photosynthesis, and lower photosynthesis in −fruit treatment was not due to water status of source leaves influenced by removing fruit. Pn tended to increase with Tl until Tl reached a critical level. Beyond the critical temperature level, Pn generally decreased. The critical Tl was roughly identified as 34–37 °C for the five cultivars. Both higher and lower substomatal CO2 (Ci) levels occurred in −fruit and −fruit* treatments than in +1 fruit and +2 fruit treatments, indicating that decreased Pn could be due to both nonstomatal and stomatal limitations. Further analysis of the relationship between Ci and photosynthetically active radiation (PAR) showed that nonstomatal limitation under low sink demand took place mostly under high PAR. Thus, high light intensity, combined with Tl may play an important role in leaf photosynthetic regulation.
The autotoxicity of root exudates and the change of rhizosphere soil microbes are two important factors that affect the quality and yield of Lanzhou lily (Lilium davidii var. unicolor). Phthalic acid (PA) is a major autotoxin of the root exudates in Lanzhou lily. In this study, we treated plants with different concentrations of PA from the Lanzhou lily root exudates and then analyzed the effects of autotoxins on fresh weight, shoot height, root length, and Oxygen Radical Absorbance Capacity in root. The diversity of soil fungi in Lanzhou lily soil was analyzed using MiSeq. The results showed that PA induced oxidative stress and oxidative damage of Lanzhou lily roots, improved the level of the membrane lipid peroxidation, reduced the content of antioxidant defense enzyme activity and the nonenzymatic antioxidant, and eventually inhibited the growth of the Lanzhou lily. We found that continuous cropping of Lanzhou lily resulted in an increase in fungal pathogens, such as Fusarium oxysporum in the soil, and reduced the size of plant-beneficial bacteria populations. The results in this study indicate that continuous cropping would damage the regular growth of Lanzhou lily.