Sesquiterpene carboxylic acids from a wild tomato species affect larval feeding behavior and survival of Helicoverpa zea and Spodoptera exigua (Lepidoptera: Noctuidae).

The sesquiterpene carboxylic acids (SCA), (+)-(E)-alpha-santalen-12-oic, (-)-(E)-endo-alpha-bergamoten-12-oic, and (+)-(E)-endo-beta-bergamoten-12-oic acid, are produced in glandular trichomes of Lycopersicon hirsutum f. typicum Humb. & Bonpl. accession (LA) 1777, which is highly resistant to a range of pests of cultivated tomatoes. L. esculentum Mill. Exposure of the larvae of two key tomato pests, tomato fruitworm [Helicoverpa zea (Boddie)] and beet armyworm [Spodoptera exigua (Hübner)], to these compounds in diets and on leaf surfaces resulted in reduced development rates and survival and deterred feeding. These effects were observed when levels of SCA, in artificial insect diet, applied to leaflets of susceptible cultivars, or synthesized in trichomes of leaves of plants, exceeded 2 mg SCA/g of diet or fresh leaf weight. This study suggests that cultivated tomatoes capable of synthesizing SCA, at 2 mg SCA/g of leaf tissue or greater, on their leaves and fruit would display enhanced host plant resistance to H. zea and S. exigua and other insect pests.

Preparative HPLC method for the purification of sulforaphane and sulforaphane nitrile from Brassica oleracea.

An extraction and preparative HPLC method has been devised to simultaneously purify sulforaphane and sulforaphane nitrile from the seed of Brassica oleracea var. italica cv. Brigadier. The seed was defatted with hexane, dried, and hydrolyzed in deionized water (1:9) for 8 h. The hydrolyzed seed meal was salted and extracted with methylene chloride. The dried residue was redissolved in a 5% acetonitrile solution and washed with excess hexane to remove nonpolar contaminants. The aqueous phase was filtered through a 0.22-microm cellulose filter and separated by HPLC using a Waters Prep Nova-Pak HR C-18 reverse-phase column. Refractive index was used to detect sulforaphane nitrile, and absorbance at 254 nm was used to detect sulforaphane. Peak identification was confirmed using gas chromatography and electron-impact mass spectrometry. Each kilogram of extracted seed yielded approximately 4.8 g of sulforaphane and 3.8 g of sulforaphane nitrile. Standard curves were developed using the purified compounds to allow quantification of sulforaphane and sulforaphane nitrile in broccoli tissue using a rapid GC method. The methodology was used to compare sulforaphane and sulforaphane nitrile content of autolyzed samples of several broccoli varieties.

Quantitative Trait Loci in Sweet Corn Associated with Partial Resistance to Stewart's Wilt, Northern Corn Leaf Blight, and Common Rust.

ABSTRACT Partial resistance to Stewart's wilt (Erwina stewartii, syn. Pantoea stewartii), northern corn leaf blight (NCLB) (Exserohilum turcicum), and common rust (Puccinia sorghi) was observed in an F(2:3) population developed from a cross between the inbred sweet corn lines IL731a and W6786. The objective of this study was to identify quantitative trait loci (QTL) associated with partial resistance using restriction fragment length polymorphic markers. Phenotypic data were collected for 2 years for Stewart's wilt, NCLB, and common rust but, due to significant family-environment interaction, analysis was conducted individually on data from each year. In 2 years of evaluation for the three diseases, a total of 33 regions in the maize genome were associated with partial resistance describing from 5.9 to 18% of the total phenotypic variability. Of six regions common in both years, three were associated with partial resistance to Stewart's wilt (chromosomes 4:07, 5:03, and 6:04), one was associated with NCLB (chromosome 9:05), and two were associated with common rust (chromosomes 2:04 and 3:04). The rust QTL on 3S mapped to within 20 cM of the rp3 locus and explained 17.7% of the phenotypic variability. Some of the QTL associated with partial resistance to the three diseases have been reported previously, and some are described here for the first time. Results suggest it may be possible to consolidate QTL from various elite backgrounds in a manner analogous to the pyramiding of major resistance genes. We also report here on two QTL associated with anthocyanin production on chromosomes 10:6 and 5:03 in the general location of the a2 gene.

Quantification of carotenoid and tocopherol antioxidants in Zea mays.

Recent investigations into carotenoid and tocopherol biological activity in mammalian systems indicate that these antioxidants are associated with the prevention of degenerative diseases. Both carotenoids and tocopherols can be found in corn kernel tissue. A replicated survey of 44 sweet and dent corn lines was conducted to determine qualitative and quantitative variability of lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, and beta-carotene, as well as the alpha-, delta-, and gamma- forms of tocopherol. The primary carotenoids in fresh market sweet corn were found to be lutein and zeaxanthin, with the gamma form dominating among the tocopherols. Mean values among the genotypes were observed to range from 0 to 20.0 and 2.4 to 63.3 microg/g dry weight for lutein and gamma-tocopherol, respectively, indicating variability among genotypes in genes regulating the metabolism of these compounds. The observed genetic variability suggests profound differences in potential health promotion among genotypes and supports the feasibility of developing germplasm with enhanced levels of these antioxidant compounds at dosages that could promote health among the consuming public.

Variation of glucosinolates in vegetable crops of Brassica oleracea.

Glucosinolates were evaluated in 5 groups and 65 accessions of Brassica oleracea (50 broccoli, 4 Brussels sprouts, 6 cabbage, 3 cauliflower, and 2 kale) grown under uniform cultural conditions. Glucosinolates and their concentrations varied among the different groups and within each group. The predominant glucosinolates in broccoli were 4-methylsulfinylbutyl glucosinolate (glucoraphanin), 3-butenyl glucosinolate (gluconapin), and 3-indolylmethyl glucosinoate (glucobrassicin). Glucoraphanin concentration in broccoli ranged from 0.8 micromol g(-1) DW in EV6-1 to 21.7 micromol g(-1) DW in Brigadier. Concentrations of the other glucosinolates in broccoli varied similarly over a wide range. In Brussels sprouts, cabbage, cauliflower, and kale, the predominant glucosinolates were sinigrin (8.9, 7.8, 9.3, and 10.4 micromol g(-1) DW, respectively) and glucobrassicin (3.2, 0.9, 1.3, and 1.2 micromol g(-1) DW, respectively). Brussels sprouts also had significant amounts of gluconapin (6.9 micromol g(-1) DW). Wide variations in glucosinolate content among genotypes suggest differences in their health-promoting properties and the opportunity for enhancement of their levels through genetic manipulation.

Carotene, tocopherol, and ascorbate contents in subspecies of Brassica oleracea.

Cruciferous vegetables contain high levels of vitamins that can act as antioxidants, compounds that may protect against several degenerative diseases. The edible portions of 50 broccoli and 13 cabbage, kale, cauliflower, and Brussels sprouts accessions were assayed to determine variation in alpha-carotene, beta-carotene, alpha-tocopherol, gamma-tocopherol, and ascorbate contents within and between subspecies of Brassica oleracea. Ascorbate content was estimated in fresh samples using HPLC. Tissues for carotene and tocopherol analysis were lyophilized prior to extraction. Carotene and tocopherol concentrations were simultaneously measured using a reverse phase HPLC system. Results indicate that there is substantial variation both within and between subspecies. Kale had the highest levels of vitamins, followed by broccoli and Brussels sprouts with intermediate levels and then by cabbage and cauliflower, with comparatively low concentrations. Variability in vitamin content among the broccoli accessions suggests that potential health benefits that accrue with consumption are genotype dependent.

Quantitative trait loci influencing chemical and sensory characteristics of eating quality in sweet corn.

This study was conducted to ascertain the chromosomal location and magnitude of effect of quantitative trait loci (QTL) associated with the chemical and sensory properties of sweet corn (Zea mays L.) eating quality. Eighty-eight RFLPs, 3 cloned genes (sh1, sh2, and dhn1), and 2 morphological markers (a2 and se1) distributed throughout the sweet corn genome were scored in 214 F2:3 families derived from a cross between the inbreds W6786su1Se1 and IL731Asu1se1. Kernel properties associated with eating quality (kernel tenderness and starch, phytoglycogen, sucrose, and dimethyl sulfide concentrations) were quantified on F2:3 sib-pollinated ears harvested at 20 days after pollination. Sensory evaluation was conducted on a subset of 103 F2:3 families to determine intensity of attributes associated with sweet corn eating quality (corn aroma, grassy aroma, sweetness, starchiness, grassy flavor, crispness, tenderness, and juiciness) and overall liking. Single factor analysis of variance revealed significant QTL for all these traits, which accounted for from 3 to 42% of the total phenotypic variation. A proportion of the RFLP markers associated with human sensory response were also found to be associated with kernel characteristics. To our knowledge this is the first report of the identification of QTL associated with human flavor preferences in any food crop. Key words : sweet corn, RFLP, quantitative trait loci, eating quality, sensory evaluation.

Identification and bioassay of kairomones forHelicoverpa zea.

Hexane extracts of leaves of 307 accessions from 73 host plant species ofHelicoverpa zea were analyzed by gas chromatography (GC) and used forH. zea oviposition and neonate larvae orientation bioassays. The gas chromatographic (GC) retention times of compounds statistically associated with behavioral activity were identified by correlation of GC peak area with oviposition and larval orientation preferences. Although taxonomically diverse in their origin, compounds for study were purified from extracts of species of the genusLycopersicon, due to their relative abundance. The structures of eight long-chain alkanes associated with oviposition preference were assigned by mass spectrometry, and the structures of five similarly associated organic acids and a terpenoid alkene were identified by(1)H and(13)C nuclear magnetic resonance spectroscopy. The structures of a number of other phytochemicals from the plant leaves were identified for comparative purposes, including a previously unknown terpene, 7-epizingiberene. Bioassays were performed on the isolated acids and on the alkane wax fractions of severalLycopersicon species, and significant differences were found in oviposition stimulation for both classes of compounds. Of the hundreds of compounds found in the extracts, none were observed to act as oviposition deterrents. The results of these bioassays may be useful in explaining the broad host range ofH. zea, as well as the process and evolution of host plant selection for oviposition.

RFLP mapping of the sugary enhancer1 gene in maize.

RFLP marker data from an F2∶3 population derived from a cross between a sugary1 (su1) and a sugary enhancer1 (su1, sel) inbred were used to construct a genetic linkage map of maize. This map includes 93 segregating marker loci distributed throughout the maize genome, providing a saturated linkage map that is suitable for linkage analysis with quantitative trait loci (QTL). This population, which has been immortalized in the form of sibbed F2∶3 families, was derived from each of the 214 F2 plants and along with probe data are available to the scientific community. QTL analysis for kernel sucrose (the primary form of sugar) concentration at 20 days after pollination (DAP) uncovered the segregation of seven major QTL influencing sucrose concentration; a locus linked to umc36a described the greatest proportion of the variation (24.7%). Since maltose concentration has previously been reported to be associated with the se1 phenotype, an analysis of probe associations with maltose concentration at 40 DAP was also conducted. The highly significant association of umc36a with maltose and sucrose concentrations provided evidence that this probe is linked to se1. Phenotypic evaluation for the se1 genotype in each F2∶3 family enabled us to map the gene 12.1 cM distal to umc36a. In contrast to previous work where se1 was reported to be located on chromosome four, our data strongly suggest that the sugary enhancer1 locus maps on the the distal portion of the long arm of chromosome 2 in the maize genome.

Genes from Lycopersicon chmielewskii affecting tomato quality during fruit ripening.

Three chromosomal segments from the wild tomato, L. chmielewskii, introgressed into the L. esculentum genome have been previously mapped to the middle and terminal regions of chromosome 7 (7M, 7T respectively), and to the terminal region of chromosome 10 (10T). The present study was designed to investigate the physiological mechanisms controlled by the 7M and 7T segments on tomato soluble solids (SS) and pH, and their genetic regulation during fruit development. The effects of 7M and 7T were studied in 64 BC2F5 backcross inbred lines (BILs) developed from a cross between LA 1501 (an L. esculentum line containing the 7M and 7T fragments from L. chmielewskii), and VF145B-7879 (a processing cultivar). BILs were classified into four homozygous genotypes with respect to the introgressed segments based on RFLP analysis, and evaluated for fruit chemical characteristics at different harvest stages. Gene(s) in the 7M fragment reduce fruit water uptake during ripening increasing pH, sugars, and SS concentration. Gene(s) in the 7T fragment were found to be associated with higher mature green fruit starch concentration and red ripe fruit weight. Comparisons between tomatoes ripened on or off the vine suggest that the physiological mechanisms influenced by the L. chmielewskii alleles are dependent on the translocation of photosynthates and water during fruit ripening.

Characterization of the effect of introgressed segments of chromosome 7 and 10 from Lycopersion chmielewskii on tomato soluble solids, pH, and yield.

Three chromosomal segments from the wild tomato L. chmielewskii have been introgressed into the L. esculentum genome. Using molecular markers they have been mapped to the middle and terminal regions of chromosome 7 (7M, 7T respectively), and to the terminal region of chromosome 10 (10T). This study was conducted to further clarify the physiological influence of the introgressed segments of chromosome 7 and 10 on tomato soluble solids (SS), and other fruit and yield parameters. The effect of the 10T segment was evaluated using five lines that differ for the presence of this segment. As previously reported this segment increased fruit pH with no significant effect on SS. Sixty-four BC2F5 backcross inbred lines (BILs) were developed from a cross using LA1501 (an L. esculentum line that contains the 7M and 7T fragments from L. chmielewskii) as the donor parent, and VF145B-7879 (a processing cultivar) as the recurrent parent. BILs were classified in four groups (+ +, inbreds without either of the L. chmielewskii segments; 7M +, lines with only the 7M segment; + 7T, inbreds with only the 7T segment, and 7M7T, inbreds with both segments) based on RFLP information, and then compared to each other for all the parameters under study. Inbreds homoyzgous for the 7M fragment displayed greater SS (26%) and higher pH (0.10) than the control group (+ +). The 7L fragment did not influence either SS or pH, but was observed to significantly increase fruit yield by 12% when compared to the recurrent parent. A gene or genes that increase yield without affecting SS or pH may have potential in the development of commerical cultivars.

Structure-activity relationships for analogs of (+)-(E)-endo-Β-bergamoten-12-oic acid, an oviposition stimulant ofHelicoverpa zea (Boddie).

Β-Bergamotenoic acid, a compound previously shown to stimulate oviposition inH. zea, was converted into a set of bicyclic analogs and tested with a set of acyclic side chain analogs to ascertain the molecular structure that maximizes insect behavioral response. While changes in the bicyclic ring elicited no variation in response, alteration in the side chain structure ofΒ-bergamotenoic acid resulted in significant changes in moth preference. Free rotation about the C-C bond proximal to the carboxylic acid group appears to be an important structural factor, since saturation of the side chain double bond significantly increased activity. The carboxylic acid group seems to be required for strong oviposition stimulation, since analogs lacking the carboxylic acid group exhibited no significant oviposition activity. Oviposition preference ofH. zea was also influenced by the length of the hydrocarbon chain to which the carboxylic acid is attached. While hexanoic acid was found inactive, the ovipositional preference for the heptanoic and octanoic acids was greatest for the one 8-carbon tested. This and other work suggest that carboxylic acids of specific chain lengths influence the oviposition behavior of bothHelicoverpa andHeliothis species and may be associated with host-plant selection. The potential use of this information in designing integrated pest management strategies for control ofH. zea is discussed.

An in vitro method for screening for the presence of thepat-2 gene in tomatoes (Lycopersicon esculentum Mill.).

Under field conditions,pat-2, the gene which conditions parthenocarpy in tomatoes, is recessive. A simple method has been devised for distinguishing the heterozygote from the two homozygotes using tissue culture. Ovaries of plants segregating for thepat-2 gene were excised and cultured on a medium containing 100 ppm gibberellic acid. After three weeks in culture, three distinct ovary sizes could be seen. It was shown, using F 3 progeny tests, that the largest ovaries corresponded to those plants homozygous for thepat-2 gene, the smallest ovaries corresponded to those plants homozygous for the wild type allele, and the intermediate sized ovaries were the heterozygotes. The ability to identify the heterozygote would greatly simplify a backcross breeding program aimed at incorporating thepat-2 gene into commercial cultivars by eliminating the need for an F 3 progeny test to determine the genotype of a plant.

Influence of trichome exudates from species ofLycopersicon on oviposition behavior ofHeliothis zea (Boddie).

Cage experiments revealed that accessions of the wild tomato speciesLycopersicon hirsutum were preferred sites for oviposition byHeliothis zea. Hexane extracts from the leaves ofL. hirsutum were also preferred sites of oviposition in choice experiments among extracts from severalLycopersicon species. Extracts ofL. hirsutum were still biologically active several days after application, indicating that the phytochemical(s) involved are relatively stable and of low volatility. Gas Chromatographic analysis of leaf hexane extracts from 12 different accessions of theL. hirsutum complex and three tomato cultivars revealed substantial qualitative and quantitative variation in the chemical composition of these extracts. Comparison of these results with extract oviposition studies implicate a group of structurally related compounds as the active agents. Mass spectroscopy has tentatively identified these compounds as sesquiterpenes with the chemical formula C15H22O2. These compounds are apparently synthesized and secreted from glandular trichomes on the leaf surface. These phytochemicals did not stimulate ovipositional behavior in females of the cabbage looper,Trichoplusia ni. The existence of genetic variation for the presence and amount of kairomones that serve as cues for insect orientation and oviposition could be utilized in a breeding program to develop tomato cultivars with genetically modified allelochemic profiles that would disrupt the sequential behavioral processes of insect host-plant selection.