A reexamination of molecular markers for use in marker-assisted breeding in tomato

Molecular markers have been used for identification and mapping of genes and QTLs for numerous agriculturally important traits in tomato, including resistance/tolerance to biotic and abiotic stresses and fruit- and flower-related characteristics. However, the extent to which markers have been utilized in tomato breeding programs has not been clearly determined. A review of the literature indicated that the utility of most markers for use in tomato breeding programs have not been verified. Many markers are not validated across tomato genotypes or are not polymorphic within tomato breeding populations. In this study, we examined the utility of available markers for several major disease resistance traits in tomato by testing them in a number of breeding lines and commercial cultivars with known resistance/susceptibility responses. While several markers were validated, others needed PCR optimization for successful amplifications or were not informative in the genotypes used. Specifically, of the 37 markers examined 19 (~51%) were informative, including markers for resistance to Fusarium wilt, late blight, bacterial wilt, tomato mosaic virus, tomato spotted wilt virus, and root knot nematodes. It appears that many of the available markers may need to be further refined or examined for trait association and presence of polymorphism in breeding lines and populations. However, with recent advances in tomato sequencing, it is becoming increasingly possible to develop more informative markers to accelerate the use of MAS in tomato breeding.     

Genotype and environmental interaction for fruit quality traits in vintage tomato varieties

Tomato (Solanum lycopersicum L.) is the second most highly consumed vegetable in the world after potato. Traits including physicochemical (lycopene, total titratable acid (TTA), total soluble solids (TSS) and vitamin C), morphological (fruit shape and size) and colors contribute to the overall fruit quality of tomato. The primary objective of the present study was to evaluate vintage tomato varieties representing a wide genetic background for fruit quality including physicochemical, morphological and color traits in multiple environments and to analyze consistency of their performances across locations. In order to achieve this objective, we acquired 44 vintage tomato varieties and evaluated them in five environments (NC, NY, OH in 2009, and NC and OH in 2010). Analysis of the data revealed that there was a significant (p < 0.01) difference among genotypes and environments for all quality traits, Genotype × Environment interaction was significant (p < 0.01) for all quality traits except for TSS. Broad-sense heritability of physicochemical traits ranged from 5.8 % (lycopene) to 35.7 % (TTA) whereas that for morphological traits ranged from 8.1 % (proximal eccentricity) to 97.3 % (fruit shape index external 1), and color from 69.0 % (a*-value) to 99.3 % (b*-value). Pearson’s correlation analysis indicated that estimated lycopene content was negatively correlated with the other physicochemical traits whereas vitamin C, TSS and TTA were positively correlated with each other. Principal component analysis (PCA) based on phenotypic data identified five components explaining at least 82 % of the total variation. Cluster analysis based on phenotypic data revealed the clusters of vintage tomato varieties were not distinct whereas single nucleotide polymorphism data revealed three distinct populations. This information including heritability and correlation coefficients from the present study may be useful to tomato breeding programs to choose germplasm, improve the response to selection and simultaneously improve multiple traits for tomato fruit quality.     

Effects of Chinese herbal medicine on plasma glucose,protein and energy metabolism in sheep

Background

The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in the past two decades and played an increasingly important role in livestock production. The present study was carried out to assess the feeding effects of a traditional nourishing Chinese herbal medicine mixture on kinetics of plasma glucose, protein and energy metabolism in sheep. Ruminal fermentation characteristics were also determined.

Methods

Four sheep were fed on either mixed hay (MH-diet) or MH-diet supplemented with 2% of Chinese herbal medicine (mixture of Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet) over two 35-day periods using a crossover design. The turnover rate of plasma glucose was measured with an isotope dilution method using [U-¹³C]glucose. The rates of plasma leucine turnover and leucine oxidation, whole body protein synthesis (WBPS) and metabolic heat production were measured using the [1-¹³C]leucine dilution and open circuit calorimetry.

Results

Body weight gain of sheep was higher (P = 0.03) for CHM-diet than for MH-diet. Rumen pH was lower (P = 0.02), concentration of rumen total volatile fatty acid tended to be higher (P = 0.05) and acetate was higher (P = 0.04) for CHM-diet than for MH-diet. Turnover rates of plasma glucose and leucine did not differ between diets. Oxidation rate of leucine tended to be higher (P = 0.06) for CHM-diet than for MH-diet, but the WBPS did not differ between diets. Metabolic heat production tended to be greater (P = 0.05) for CHM-diet than for MH-diet.

Conclusions

The sheep fed on CHM-diet had a higher body weight gain and showed positive impacts on rumen fermentation and energy metabolism without resulting in any adverse response. Therefore, these results suggested that the Chinese herbal medicine mixture should be considered as a potential feed additive for sheep.     

Novel molecular marker associated with Tm2a gene conferring resistance to tomato mosaic virus in tomato

Tomato mosaic virus (ToMV) is an important Tobamovirus that causes significant crop losses. Resistance to the ToMV is conferred by the genes Tm1, Tm2 and Tm2^a. Among these three genes, Tm2^a confers resistance to most strains of the ToMV. Screening of genetic lines under field conditions based on phenotype is time‐consuming and challenging due to concerns associated with stability of the virus and its potential transmission to other plants. Tightly linked molecular markers associated with resistance genes can improve selection efficiency and avoid these problems. This study developed a PCR‐based marker based on restriction site differences from Tm2^a locus‐specific sequences, which was found to be useful in identifying the resistant and susceptible genotypes and was consistent with phenotypic data. The marker is a codominant cleaved amplified polymorphic sequence (CAPS) marker producing 270‐ and 600‐bp DNA fragments from resistant genotypes and an 870‐bp fragment from susceptible genotypes when digested with HaeIII restriction enzyme. This novel marker can be useful for tomato breeders to screen progeny from segregating populations for ToMV resistance.     

Resistance to Xanthomonas perforans race T4 causing bacterial spot in tomato breeding lines

Tomato (Solanum lycopersicum) is the second most important vegetable crop in the world. Bacterial spot (BS) of tomato, caused by four species of Xanthomonas: X. euvesicatoria, X. vesicatoria, X. perforans and X. gardneri, results in severe loss in yield and quality due to defoliation and formation of lesions on fruits, respectively. Currently management practices do not offer effective control under conditions of high disease pressure. Thus, developing BS resistance is a critical priority for tomato growers in order to minimize crop losses. Sixty‐three advanced tomato breeding lines, heirlooms and wild tomato lines with diverse genetic backgrounds were screened under greenhouse and field conditions for BS resistance using X. perforans race T4, which was found to be a prevalent race in North Carolina. Race T4 isolate 9 was used to inoculate the plants by spraying, and disease severity was measured using the Horsfall–Barratt scale. Tomato lines 74L‐1W(2008), NC2CELBR, 081‐12‐1X‐gsms, NC22L‐1 (2008) and 52LB‐1 showed resistance to BS in the field and/or greenhouse trials. These lines were derived from S. pimpinellifolium L3707. Screening L3707 followed by development of a mapping population and mapping resistance genes might be useful for breeding resistance against BS in future breeding programmes.     

Quantitative trait loci for agronomic traits in soybean

There continues to be improvement in seed yields of soybean by conventional breeding, but molecular techniques may provide faster genetic gains. The objective of this study was to identify quantitative trait loci (QTL) associated with the agronomic traits seed yield, lodging, plant height, seed filling period and plant maturity in soybean. To achieve this objective, 101 F₆‐derived recombinant inbred lines (RIL) from a population developed from a cross of N87‐984‐16 × TN93‐99 were used. Experiments were conducted in six environments during 2002–2003. Heritability estimates on an entry mean basis from data combined across environments ranged from 0.12 to 0.65 for seed yield and seed filling period, respectively. Composite interval mapping detected one QTL for yield (near Satt076), two for lodging (near Satt225 and Satt593) and four for maturity (near Satt263, Satt292, Satt293 and Satt591) in this population. Additional environmentally sensitive QTL for these traits, and for seed filling period and plant height are also reported. The QTL associated with agronomic traits that we report and the recently released germplasm (PI 636460) from this population may be useful in soybean breeding programmes.     

Effects of Chinese herbal medicine on plasma glucose,protein and energy metabolism in sheep

Background： The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in the past two decades and played an increasingly important role in livestock production. The present study was carried out to assess the feeding effects of a traditional nourishing Chinese herbal medicine mixture on kinetics of plasma glucose, protein and energy metabolism in sheep. Ruminal fermentation characteristics were also determined. Methods： Four sheep were fed on either mixed hay （MH-diet） or MH-diet supplemented with 2% of Chinese herbal medicine （mixture of Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet） over two 3S-day periods using a crossover design. The turnover rate of plasma glucose was measured with an isotope dilution method using [U-^13C]glucose. The rates of plasma leucine turnover and leucine oxidation, whole body protein synthesis （WBPS） and metabolic heat production were measured using the [1-^13C]leucine dilution and open circuit calorimetry. Results： Body weight gain of sheep was higher （P = 0.03） for CHM-diet than for MH-diet. Rumen pH was lower （P = 0.02）, concentration of rumen total volatile fatty acid tended to be higher （P = 0.05） and acetate was higher （P = 0.04） for CHM-diet than for MH-diet. Turnover rates of plasma glucose and leucine did not differ between diets. Oxidation rate of leucine tended to be higher （P = 0.06） for CHM-diet than for MH-diet, but the WBPS did not differ between diets. Metabolic heat production tended to be greater （P = 0.05） for CHM-diet than for MH-diet. Conclusions： The sheep fed on CHM-diet had a higher body weight gain and showed positive impacts on rumen fermentation and energy metabolism without resulting in any adverse response. Therefore, these results suggested that the Chinese herbal medicine mixture should be considered as a potential feed additive for sheep.     

Diversity Analysis of Garlic (Allium sativum L.) Germplasms Available in Nepal Based on Morphological Characters

A total of 179 garlic (Allium sativum L.) accessions were collected from various parts of Nepal in 2000. Each accession was planted in each of a plot of 2.25 m² at the Institute of Agriculture and Animal Science (IAAS) Horticulture farm (225 m asl) and at the Agriculture Research Station, Dailekh (1400 m asl) of Nepal Agriculture Research Council (NARC) in the first week of November 2000. All accessions were characterized for leaf erectness, leaf color, leaf wax, leaf cross section, bulb regularity, bulb skin color, bulb outer scales number, days to emergence, bulbing period, number of green leaves at 135 days after planting, days to maturity, plant height, bulb weight, bulb diameter, number of cloves per bulb, clove diameter and bulb yield. Data were analyzed by using principal component and cluster analysis procedures to reveal three major clusters. Four principal components were identified explaining more than 86% of total variation. Major characters included in the principal components were bulb weight, diameter, yield, number of cloves per bulb, maturity, plant height, number of green leaves at 135 days after planting, and bulbing period. The level of variation found in the collection showed the great potentiality of improving agronomic characters in garlic.     

Modifier QTL for fatty acid composition in soybean oil

Soybean [Glycine max (L.) Merr.] is the principal oilseed crop in the world. Soybean oil has various industrial and food applications. The quality of soybean oil is determined by its fatty acid composition. Palmitic, stearic, oleic, linoleic and linolenic are the predominant fatty acids in soybean oil. The objective of this study was to determine the associations of simple sequence repeat (SSR) molecular markers with minor differences in fatty acids in soybean oil thereby detecting modifier quantitative trait loci (QTL) which could further improve soybean oil quality. To achieve this objective, 101 F₆-derived recombinant inbred lines (RIL) from a population whose parents did not contain major mutant fatty acid alleles were developed from a cross of N87-984-16 × TN93-99. Fatty acids were determined by gas chromatography. Heritability estimates on an entry mean basis for fatty acids ranged from 65.8 to 77.3% for palmitic and linoleic acids, respectively. Molecular marker Satt537 located on molecular linkage group (MLG) D1b was associated with palmitic acid and Satt168 and Satt249 located on MLG B2 and J, respectively were associated with stearic acid. Molecular markers Satt185 or Satt268 (which are within 0.6 cM of each other) located on MLG E were consistently associated with oleic and linoleic acid, and Satt263 and Satt235 located on MLG E and G, respectively were associated with linolenic acid. The lack of markers associated with multiple fatty acids suggests the possibility of independently changing fatty acid levels to achieve a desirable composition, except for regions common to all saturated fatty acids. Phenotypic variation explained by the fatty acids modifier QTL ranged from 10 to 22.5%. These modifier QTL may be useful in making minor improvements to further enhance the quality of soybean oil.