Carotenoids are isoprenoid compounds synthesized in plants that serve as photoprotectants essential for photosynthesis and provide plant tissues with red, orange, and yellow pigmentation. These compounds are important in human health, because they serve as both vitamin A precursors as well as having antioxidant properties. Carrot (Daucus carota ssp. sativus) provides an important source of carotenoids in the human diet, providing up to 30% of provitamin A in the United States. Although essential to human health, very little is currently understood about the accumulation of carotenoids in carrot. To better understand the molecular mechanism for carotenoid accumulation in carrot, we used reverse-transcription quantitative polymerase chain reaction (PCR) to evaluate the expression of nine genes in the carotenoid biosynthetic pathway in storage root tissue. No significant difference was found among white, yellow, orange, and dark orange carrot roots in seven of the nine genes evaluated. However, increased phytoene synthase 1 (PSY1) and phytoene synthase 2 (PSY2) expression was observed in orange and dark orange carrot roots compared with yellow and white carrots. Increased PSY1 and PSY2 expression was not observed in the leaf tissue of these genotypes, indicating a different mechanism for carotenoid accumulation in the leaf tissue of carrot. This study is the first to demonstrate that naturally occurring mutations that dramatically increase carotenoid accumulation in orange carrot are associated with increased PSY1 and PSY2 expression and it provides insights into the mechanism underlying the biosynthesis of these important photoprotectants and nutrients.
Megan J. Bowman, David K. Willis, and Philipp W. Simon
Jack E. Staub, Vanessa S. Gordon, Philipp Simon, and Todd C. Wehner
J. Michele Myers, Philipp W. Simon, M.E.N. Fonseca, and Leonardo S. Boiteux
Garlic is an asexually propagated crop in which the greatest yield losses are attributed to virus infection. Currently, virus-free garlic is produced through shoot tip culture, and there are no known naturally occurring resistant clones. This study evaluated garlic germplasm (propagated from typical bulbs, not shoot tips) for incidence of two viruses known to infect garlic (onion yellow dwarf virus, OYDV and leek yellow stripe virus, LYSV) using dot blot ELISA. Young leaf tissue was collected from 173 garlic clones. For 118 clones, plants grown in the field from typical bulbs only were evaluated. For 55 clones, plants grown in the greenhouse from both bulbs and topsets (bulbils) were evaluated. Topsets are small bulbs that are produced in the inflorescence of stalking garlic. Each clone was tested at least three times for incidence of both viruses. In field grown bulbs, we found that 70% were infected with OYDV and 85 % were infected with LYSV. In greenhouse grown samples, incidence of OYDV was generally higher in plants from topsets than those from bulbs while no differences were seen for LYSV. Three clones were negative for both viruses and might be a useful source of resistance that can be used in producing virus resistant lines.
Aneela Nijabat, Adam Bolton, Muhammad Mahmood-ur-Rehman, Adeel Ijaz Shah, Rameez Hussain, Naima Huma Naveed, Aamir Ali, and Philipp Simon
Heat waves occur with more regularity and they adversely affect the yield of cool season crops including carrot (Daucus carota L.). Heat stress influences various biochemical and physiological processes including cell membrane permeability. Ion leakage and increase in cell permeability are indicators of cell membrane stability and have been used to evaluate the stress tolerance response in numerous crops and inform plant breeders for improving heat tolerance. No study has been published about the effects of heat stress on cell membrane stability and relative cell injury of carrot. Therefore, the present study was designed to estimate these stress indicators in response to heat stress at the early and late seedling developmental stages of 215 diverse accessions of wild and cultivated carrot germplasm. The article identifies the relationship between early and late stages of seedling tolerance across carrot genotypes and identifies heat-tolerant genotypes for further genetic analysis. Significant genetic variation among these stress indicators was identified with cell membrane stability and relative cell injury ranging from 6.3% to 97.3% and 2.8% to 76.6% at the early seedling stage, respectively; whereas cell membrane stability and relative cell injury ranged from 2.0% to 94.0% and 2.5% to 78.5%, respectively, at the late seedling stage under heat stress. Broad-sense heritability ranged from 0.64 to 0.91 for traits of interest under study, which indicates a relatively strong contribution of genetic factors in phenotypic variation among accessions. Heat tolerance varied widely among both wild and cultivated accessions, but the incidence of tolerance was higher in cultivated carrots than in wild carrots. The cultivated carrot accessions PI 326009 (Uzbekistan), PI 451754 (Netherlands), L2450 (USA), and PI 502654 (Pakistan) were identified as the most heat-tolerant accessions with highest cell membrane stability. This is the first evaluation of cell membrane stability and relative cell injury in response to heat stress during carrot development.
James M. Bradeen, Inga C. Bach, Mathilde Briard, Valérie le Clerc, Dariusz Grzebelus, Douglas A. Senalik, and Philipp W. Simon
A sample of 124 Daucus carota L. accessions, including cultivated carrot [D. carota ssp. sativus (Hoffm.) Arcangeli] and related wild subspecies, using a variety of molecular markers was examined. Represented within the samples were wild accessions from 18 countries, 14 of 16 major root types of European origin, and examples of major North American and Asian cultivated carrot types. Amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) markers revealed extensive variation within D. carota. Although cultivated carrot and wild D. carota subspecies can cross freely, cultivated and wild carrots clustered separately, supporting the possibility that human selection for desirable horticultural traits has artificially reduced gene flow between cultivated and wild forms. Our analyses support the likelihood that North American D. carota populations arose due to introduction of weedy materials rather than escape of cultivated forms. With the exception of wild vs. cultivated types, no genetic alliances were evident in dendrogram topology. Furthermore, between and even within nonmapped marker classes, dendrogram topology predictions were not consistent. Generally poor correlations among root types, geographic origin, mitochondrial, plastid, and specific nuclear diversity and AFLP/ISSR data were also observed. We concluded that genetic diversity in carrot is extensive and relatively nonstructured in nature.
Adam Bolton, Aneela Nijabat, Muhammad Mahmood-ur-Rehman, Naima Huma Naveed, A.T.M. Majharul Mannan, Aamir Ali, Mohamed A. Rahim, and Philipp Simon
Carrot production is constrained by high levels of heat stress during the germination stage in many global regions. Few studies have been published evaluating the effect of heat stress on carrot seed germination or screening for genetic heat stress tolerance. The objectives of this study were to evaluate the response of diverse carrot germplasm to heat stress, identify heat-tolerant germplasm that may be used by plant breeders, and define the appropriate temperature for assessing heat tolerance in germinating carrot seed. To identify an appropriate screening temperature, three commercial hybrids and an open pollinated variety were evaluated at five temperatures (24, 32.5, 35, 37.5, and 40 °C). In preliminary studies, 35 °C was identified as the optimal temperature for screening heat tolerance of carrot seed. Cultivated and wild carrot plant introductions (PIs) (n = 270) from the U.S. Department of Agriculture (USDA) National Plant Germplasm System (NPGS) representing 41 countries, inbred lines from the USDA Agricultural Research Service (n = 15), and widely grown commercial hybrids (n = 8) were evaluated for heat tolerance under heat stress and nonstress conditions (35 °C and 24 °C, respectively) by calculating absolute decrease in percent germination (AD), inhibition index (II), relative heat tolerance (RHT), and heat tolerance index (HTI). All measurements of heat tolerance identified significant differences among accessions; AD ranged from −13.0% to 86.7%, II ranged from 35.7% to 100.0%, RHT ranged from 0 to 1.36, and HTI ranged from 0.0 to 1.45. The broad-sense heritability (H2) calculations ranged from 0.64 to 0.86 for different traits, indicating a moderately strong genetic contribution to the phenotypic variation. Several wild carrot accessions and inbred lines displayed low levels of heat tolerance, whereas cultivated accessions PI 643114 (United States), PI 652400 and PI 652403 (Turkey), PI 652208 (China), and PI 652403 (Russia) were most heat tolerant. This is the first evaluation of heritability for heat stress tolerance during carrot seed germination, the first measure of HTI, and the first correlation calculation between heat and salt tolerance during germination in carrot.