To understand the prospects of applying the RAPD technique to assay genetic diversity in Ipomoea, four species (I. batatas, I. trifida, I. triloba, and I. ×leucantha) were analyzed for RAPD molecular markers. Six accessions of each species were used. Significant RAPD polymorphisms were detected within each species. Of 20 primers used, nine produced clear scorable polymorphic bands. The number of polymorphic bands produced per primer ranged from two to nine. Pair-wide genetic distance was calculated based on “band sharing”. The SAS-CLUSTER procedure was used to build a hierarchical species dendrogram. The four species were clearly separated by the clustering, which agrees with their existing taxonomic relationship. This study shows that RAPD analysis can be a powerful tool for identifying duplicates of germplasm acessions and for assessing genetic diversity. The procedures are relatively inexpensive and easy to perform and could be valuable in preliminary assessment of field genebank collections to separate species and indicate duplications in collected material.
Dapeng Zhang and Wanda Collins
Dapeng Zhang and Wanda W. Collins
Randomly amplified polymorphic DNA (RAPD) analysis was performed on 18 accessions belonging to four different species of the genus Ipomoea, including sweetpotato and three related species. Twenty-two out of 30 primers tested revealed polymorphisms among these four species. Eight primers were selected on the basis of the number and repeatability of polymorphism produced. With these, a total of 98 different DNA bands were obtained and 85% of them were polymorphic. Based on the presence/absence of the bands, a genetic similarity among accessions and among species was calculated. Unweighted pair-group method with arithmetical averages (UPGMA) based on the similarity coefficients clearly discriminated these four species. Ipomoea trifida and sweetpotato share more genetic similarity. Ipomoea triloba and I. leucantha fall into another cluster. This study demonstrated that RAPD techniques can be a very useful tool for genotype/accession identification and studying the genetic relationship among genotypes/accessions of sweetpotato and among species of Ipomoea.
Dapeng Zhang, Wanda W. Collins, and Maria Andrade
Sweetpotato [Ipomoea batatas (L.) Lam.] is intensively used as an animal feed in many developing countries. Information about trypsin inhibitor activity (TIA), an antinutritional component in this crop, will be useful for breeding sweetpotato as animal feed. Nine sweetpotato lines were grown at two locations and fertilized or nonfertilized conditions at each location. Samples were analyzed for TIA using a substrate-specific colorimetric method. Soybean [Glycine max (L.) Merr.] seeds were used to compare the levels of TIA in sweetpotato and soybean. Activity in roots ranged from 29.5 to 55.0 units in the nine lines. The mean TIA in roots was 40.7 units averaged over lines and environments, which was ≈28% of the mean for the five soybean cultivars. Activity in sweetpotato vines was only ≈14.6% of that in the roots, and TIA in fertilized plots was 150% and 67% higher than that in nonfertilized plots in the two locations, respectively. There was a small but significant positive correlation between TIA and crude protein in roots. These results suggested that TIA in sweetpotato storage roots may be high enough to have a substantial nutritional impact on animals, whereas TIA in vines is very low and should be of less nutritional concern.
Dapeng Zhang, Wanda W. Collins, and Suzanne Belding
Eight sweetpotato [Ipomoea batatas (L.) Lam.] clones were evaluated for the digestibility of their starch in animals with a simple in vitro screening method. Starch digestibility varied significantly among clones. After dry-heat treatment at 100C for 30 minutes, digestibility of the most heat-sensitive clone increased only 37.8%. Excellent repeatable results were obtained with a simple weight-loss method. This assay procedure can be used as a screening method in breeding digestible sweetpotatoes for animal feed.
Dapeng Zhang, Wanda W. Collins, and Maria Andrade
Two experiments that included 25 sweetpotato [Ipomoea batatas (L.) Lam] genotypes were planted in various environments across North Carolina, and an in vitro screening method was used to investigate genotypic and environmental variance and genotype × environment (G × E) interactions of starch digestibility in sweetpotato. Significant genotypic variation of starch digestibility was found in both experiments. Some clones have starch digestibility equivalent to that of corn. Variance analysis from both experiments indicated that genotypic variance was the dominant component in starch digestibility. G × E interaction only accounted for 6.8% of the phenotypic variance in one experiment and for 5.9% in the other one. These results suggested that starch digestibility of sweetpotato could be improved to a level as that of corn through conventional breeding.
Da-Peng Zhang, Zi-Lian Zhang, Jia Chen, and Jiang Lu
The abscisic acid (ABA) has a key role in the regulation of grapevine fruit ripening, but the cellular and molecular biological mechanism of the hormone action in the fruit ripening remains unknown. By means of differential centrifugation, microsomes were prepared from Kyoho grapevine (Vitis vinifera L. × V. Labrusca L.) berries, and using the microsomes, we have obtained evidence for the occurrence of specific ABA-binding sites on the membranes with the microvolume radio-ligand binding assay. The binding sites had saturability, high affinity, and low capacity. The results of trypsin and dithiothreitol treatments to the microsomes suggested that ABA binding sites had the properties of proteins that might have disulfide group located at or near the binding site. The binding maximum amount of ABA in the microsomes was at pH 6.0 and the activity of ABA binding proteins was higher at 25 than at 0°C (temperature). The amount of ABA bound reached 54% of the ABA binding maximum (Bmax) for 10 minutes of incubation and Bmax reached for 30 minutes. The dissociation constant (Ka) and Bmax of ABA binding proteins in the microsomes were 17.5 nmol/L and 98.4 fmol/mg protein, respectively.
Da-Peng Zhang, Zi-Lian Zhang, Jia Chen, and Jiang Lu
By using the micro-volume radio-ligand binding essay, the changes in the kinetic characteristics of the abscisic acid (ABA)-binding protein(s) of the Kyhoh grapevine (Vitis vinifera × V. labrusca) fruit during the different stages of fruit development have been studied. The changes in the berry volume growth, concentration of sugar, organic acids, and ABA in fruit mesocarp have been surveyed, especially for studies of ABA-binding protein. The dissociation constant (Kd) and ABA binding maximum (Bmax) were determined by the Scatchard plots for ABA binding in microsomes of the fruit. They are Kd = 17.5, 50.0, 6.3, 13.3 nmol·L–1; Bmax = 98.6, 523.0, 41.6, 85.4 μmol·mg–1 protein, respectively, for the fruit developmental phase I, II, veraison, and phase III. The Scatchard plots showed a rectilinear function for all of the developmental phases including veraison, which suggests the sole ABA-binding site of high affinity for ABA in the fruit microsomes, but this site could either be only one kind of the same protein or consist of more kinds of different proteins for different developmental stages. The binding affinity of ABA-binding protein(s) for ABA was shown to be higher at veraison time than during other developmental phases; this binding affinity increased nearly by 10 times from phase II to veraison, while the concentration (Bmax) of the ABA-binding protein(s) decreased to the minimum at veraison. The very low concentration of ABA at veraison may be able to trigger the onset of fruit ripening due to the increase of the binding affinity of ABA-binding protein(s) for ABA at this time. The possible functions of the ABA-binding protein(s) for fruit development during the different developmental stages were discussed, and it is suggested that the protein(s) detected could be the putative ABA receptor(s) or transporter(s) for the action of this plant hormone in grapevine.
Zimian Niu*, Dapeng Zhang, Hongyu Zhao, and Curt Rom
The volatile aromas from the fruits of `Naganofuji No.2' apple (Malus domestica Mill.) were determined by gas chromatography (GC) and combined GC-mass spectrometry (GC-MS) after different temperature conditions. The fruits from CA storage were sealed in glass and the volatiles in the headspace were determined. Eleven compounds of four chemical classes from active carbon absorbed samples were measured and three of them—tormic acid pentyl ester, butanoic acid-1-methyl ethylester and 4-hydroxy-3-methyl-2-butanone, were identified at 20 °C, but not at °C. Under 20 °C condition, the contents of three volatiles increased from 1 hour and reached to their peaks at the 4th to 7th hour. The content of ethylene reached its peak at 4 hours and changed synchronically with the other volatiles during the experiment. The content of ethylene was significantly positively correlated with the contents of volatile aromas (r = 0.96-0.98, P ≤ 0.01). Under °C condition, the content of ethylene was significant lower than that of at 20°C and there was no ethylene peak produced during experiment. When the fruits were treated with ethephon (0.1 mg·L-1) at 5°C, the content of ethylene increased greatly. The highest level of ethylene was found at 4 to 7 hours and the peaks of volatiles also appeared at 7 hours or 10 hours after the treatment. It was suggested that the production of ethylene in fruits could be thought as an indicator of some volatile aromas.
Zimian Niu, Dapeng Zhang, Jicheng Zhan, and Curt Rom
Influence of photosynthetically active radiation (PAR) microclimate in the canopy of four training systems [open-center with high trunk (OH); open-center with middle trunk (OM); open-center with low trunk (OL); and a traditional round canopy (RC)] on the growth, yield and fruit quality of apple were studied in the Beijing area. The results showed that: 1) the growth and yield potential were affected by canopy light microclimate. The average leaf chlorophyll content from OH, OM and OL systems was 12.3% to 18.1% greater than that from the RC system. Trees from OH, OM, and OL systems produced 84.2% to 89.7% of shoot forming flower clusters compared to only 47.3% to 50.9% of the RC shoots. Training system did not affect total yield of 8-year-old trees, but in 10-year-old trees the RC system had lower yields compared with open-center systems. 2) Fruit quality was also affected by canopy light microclimate. The average anthocyanin content in the skin of fruit from OH, OM, and OL systems was 35.9% to 46.1% higher than that from the RC system, but chlorophyll content from the OL system was higher than in the open-center systems. Meanwhile, the contents of TSS and esters in apple flesh from the open-center systems were significantly higher than that from the RC system. 3) When the relative value of PAR in canopy exceeded 33.8%, the growth index of trees (chlorophyll: return-bloom ratio) exceeded 66.6% and the fruit quality index (TSS × anthocyanin) exceeded 94.7%. When PAR was less than 20.6%, the growth index was under 37.2% and the fruit quality index was under 67.5%. PAR value was significantly correlated with the growth and fruit quality index in the four training systems, and the total canopy volume of higher PAR(exceeding 33.8%, relative value) from OH, OM, and OL systems was 37.1% to 45.0% greater than that from the RC system.
Viji Sitther, Dapeng Zhang, Sadanand A. Dhekney, Donna L. Harris, Anand K. Yadav, and William R. Okie
Information on genetic relationships and pedigree structure in germplasm collections is vital to breeders in crop improvement programs. In this study, we assessed genetic identity, kinship distance, and parentage–sibship relationships among 37 peach (Prunus persica) accessions and breeding lines using simple sequence repeat (SSR) markers. Pairwise comparisons based on multilocus SSR profiles led to the identification of two synonymous groups including five accessions. Two pairs of parent–offspring and one full sibling relationships were identified using the likelihood method, and Bayesian cluster analysis partitioned the accessions into groups that were partially compatible with the known pedigree, origin, and flesh color. The 37 accessions were grouped into four clusters, which were largely supported by the known pedigree and origin of these accessions. Although the observed mean heterozygosity was 0.219, mean inbreeding coefficient was 0.635, indicating a high degree of inbreeding among the accessions. Eleven of the 15 SSR markers (73.3%) tested were transferable to nine related Prunus species. Results of the study demonstrate that these SSRs could facilitate the assessment of genetic identity and pedigree structure.