Using two sweetpotato (Ipomoea batatas (L.) Lam) F1 populations from diverse environments we investigated the AFLP marker profiles of the genotypes for association studies between the molecular markers and southern root-knot nematode (Meloidogyne incognita) resistance expression. Population one consisted of 51 half-sib genotypes developed at the Louisiana State Univ. AgCenter. The second population consisted of 51 full-sibs developed by the East African and International Potato Center sweetpotato breeding programs. Results for nematode resistance expression indicate a binomial distribution among the genotypes. Using analysis of molecular variance, logistic regression and discriminant analysis, AFLP markers that are most influential with respect to the phenotypic trait expression were selected for both populations. A comparative analysis of the power of models from the two statistical models for southern root-knot nematode resistance class prediction was also done. The diversity and possible universal similarity of influential markers between the two populations and the expected impact in sweetpotato breeding programs will be discussed.
Mwamburi Mcharo*, Don Labonte, Chris Clark, and Mary Hoy
Amanda Garris, Lindsay Clark, Chris Owens, Steven McKay, James Luby, Kathy Mathiason, and Anne Fennell
In grapevines (Vitis spp.), the timing of growth cessation in the fall is an important aspect of adaptation and a key objective in breeding new grape cultivars suitable for continental climates. Growth cessation is a complex biological process that is initiated by environmental cues such as daylength and temperature, as well as water and nutrient availability. The genetic control of growth cessation in grapevines was studied by mapping quantitative trait loci (QTL) in a hybrid grape population. An F2 mapping population was developed by selfing a single F1 plant derived from a cross between an accession of the North American species Vitis riparia and the Vitis hybrid wine cultivar Seyval (Seyve-Villard 5–276). A linkage map was constructed using 115 simple sequence repeat (SSR) markers and six candidate genes in a population of 119 F2 progeny. The markers provided coverage of the 19 Vitis linkage groups with an average distance between markers of 8.4 cM. The critical photoperiod for growth cessation in lateral buds for the parents and F2 progeny was determined in a replicated field trial in 2001 and 2002 and under controlled photoperiod treatments in a greenhouse in 2002, 2003, and 2004. QTL analysis using composite interval mapping identified a single major QTL in the field and greenhouse trials. However, the field and greenhouse QTL mapped to different linkage groups in the two different environments, suggesting the presence of additional, nonphotoperiodic cues for induction of growth cessation in the field. In the greenhouse, where noninducing temperatures were maintained, a QTL on linkage group (LG) 13 explained 80.0% to 96.6% of the phenotypic variance of critical photoperiod for growth cessation. In the field, where vines experienced natural fluctuations in temperature and rainfall in addition to the naturally decreasing photoperiod, a QTL on LG 11 explained 85.4% to 94.3% of phenotypic variance.
David G. Clark, Chris Dervinis, Francine Cuquel, Harry Klee, Jim Barrett, and Terril Nell
In an effort to modify and study leaf senescence, we have produced several different transgenic petunia lines with altered leaf senescence phenotypes. Using two promoters from senescence-associated genes (sag12 & sag13) fused to the isopentenyl transferase (IPT) gene, which catalyzes the rate-limiting step of cytokinin production, we have produced transgenic petunia plants with delayed lower leaf senescence. We have observed that apparent “leaky” expression of IPT gives rise to plants with other morphological alterations such as increased branching habit and decreased root formation. Plants with delayed leaf senescence phenotypes were selected and bred to produce progeny that were evaluated in greenhouse experiments. Breeding characteristics, horticultural performance and reproduction of these plants will be discussed in terms of potential commercial benefits and limitations. Using the sag12 promoter to drive expression of the knotted (KN1) gene, we have also been able to engineer petunia plants with delayed lower leaf senescence. Initial progeny evaluations of sag12-KN1 petunias will also be discussed.
David G. Clark, Chris Dervinis, James E. Barrett, Harry Klee, and Michelle Jones
Cytokinins have been shown to delay the onset of leaf senescence. The focus of this project was to produce transgenic petunia (Petunia ×hybrida) plants that over-produced endogenous cytokinins in a senescence specific manner. This was achieved by transforming plants with the IPT (isopentenyl transferase) gene driven by the senescence-associated transcriptional promoter, PSAG12. Two independent transgenic events produced T1 and T2 generation seedling lines that demonstrated the desired nonsenescent phenotype in progeny trials. These lines were used to evaluate the horticultural performance of PSAG12-IPT petunia plants in terms of delayed senescence, rooting of vegetative cuttings, lateral branch growth, flower number, floral timing, and fruit set. Although both lines displayed a delayed senescence phenotype the two PSAG12-IPT transgenic lines differed from each other in regard to other horticultural traits. In addition to delayed leaf senescence, line I-1-7 also demonstrated a decrease in adventitious rooting and an increased number of branches during plant production. Line I-3-18 also demonstrated a delayed leaf senescence phenotype; however, plants of this line were not greatly altered in any other horticultural performance traits in comparison to wild-type `V26'. IPT transcript was detected in young fully expanded leaves of both lines, although mRNA levels were higher in I-1-7 plants. A greater than 50-fold increase in IPT transcript abundance was detected in leaves of transgenic plants following drought stress. These results demonstrate that it is possible to use PSAG12-IPT to produce transgenic plants with delayed leaf senescence but differences in plant morphology between transgenic lines exist, which may alter horticultural performance characteristics.
Jinquan Feng, Andrew V. McGlone, Mike Currie, Chris J. Clark, and Bob R. Jordan
Dry matter content (DM), soluble solids content (SSC), and flesh hue (FH) are important quality attributes of yellow-fleshed kiwifruit (Actinidia chinensis ‘Hort16A’). Rapid non-destructive measurement tools enable effective assessment of quality attributes in the field or in packhouses and offer the potential for selective harvesting and differential postharvest treatments. This study investigates a few practical issues for use of one such tool, a portable near-infrared spectrometer, the FANTEC™ FQA-NIRGUN. Experimental results indicated that the precision of the NIRGUN in predicting the DM, SSC and FH of ‘Hort16A’ kiwifruit, given as the root mean square error of calibration (RMSEC), is 0.6%, 0.9% and 1.4°, respectively, and is similar to that reported for standard laboratory benchtop spectrometers. These levels of precision are good enough to distinguish differences in fruit quality between fruit from different vines. However, this is only possible when measurements are carried out under similar conditions either within a short period of time in the field when fruit temperature is stable or in a temperature-controlled laboratory. The calibration developed on one instrument could be transferred to another instrument, but bias correction is necessary after the calibration transfer. Hence, the absolute accuracy of predictions from the NIRGUN is limited by significant and variable bias offset issues, particularly for field measurements when fruit are on the vine.
Cecilia E. McGregor, Douglas W. Miano, Don R. LaBonte, Mary Hoy, Chris A. Clark, and Guilherme J.M. Rosa
Sweet potato virus disease (SPVD) is one of the most devastating diseases affecting sweetpotato (Ipomoea batatas), an important food crop in developing countries. SPVD develops when sweetpotato plants are dually infected with sweet potato feathery mottle virus (SPFMV) and sweet potato chlorotic stunt virus (SPCSV). To better understand the synergistic interaction between these viruses, global gene expression was previously studied in the susceptible cultivar Beauregard. In the current study, global gene expression between SPVD-affected plants and virus-tested control plants (VT) were compared in ‘Beauregard’ (Bx) and resistant ‘NASPOT 1’ (Nas) sweetpotato cultivars at 5, 9, 13, and 17 days post inoculation (DPI). Titer levels of SPFMV and SPCSV were significantly lower in inoculated resistant plants (Nas_SPVD) than in susceptible plants (Bx_SPVD) at most of the time points. Chloroplast genes and cell expansion-related genes (including xyloglucan endotransglucosylase/hydrolases) were suppressed in Bx_SPVD, while stress-related genes were induced. This trend was not observed in resistant NAS_SPVD. Genes related to protein synthesis (e.g., ribosomal proteins and elongation factor genes) were induced in resistant NAS_SPVD at 5 DPI before returning to levels comparable with NAS_VT plants. At this time (5 DPI), individual viruses could not be detected in NAS_SPVD samples, and no symptoms were observed. Induction of protein synthesis-related genes is common in susceptible plants after virus infection and is generally in proportion to virus accumulation. Our results show that induction of protein synthesis genes also occurs early in the infection process in resistant plants, while virus titers were below the level of detection, suggesting that virus accumulation is not required for induction.