scholarly journals Identities of and Phenotypic Variation for Epicuticular Waxes among Leaves and Plants from Inbred Onion Populations

HortScience ◽  
2020 ◽  
Vol 55 (12) ◽  
pp. 2008-2010
Author(s):  
Jiffinvir Khosa ◽  
Derek Hunsaker ◽  
Michael J. Havey

The amounts and types of epicuticular waxes on onion (Allium cepa) leaves affect feeding damage by onion thrips (Thrips tabaci). This study used gas chromatography mass spectrometry (GCMS) to establish the identities of waxes and measure over time wax amounts on leaves of inbred onion plants with glossy, semiglossy, and waxy foliage. Nine waxes were detected on leaves of all inbreds, and higher coefficients of variation (cv) were observed for less abundant waxes on foliage of doubled haploid onions. Older leaves had higher amounts of waxes compared with younger leaves on the same plant. Except for one minor wax, amounts of individual waxes on leaves were not significantly different for plants of different ages. There was a significant inbred by sampling date interaction due to lower amounts of waxes on the leaves of older plants from the semiglossy inbred. These results indicate that there is little advantage to multiple samplings of leaves over time from the same plant and resources may be better used to evaluate more plants. The relatively large cvs for amounts of specific waxes may reduce response to selection for unique epicuticular wax profiles to develop onion populations that suffer less feeding damage by onion thrips.

2014 ◽  
Vol 139 (4) ◽  
pp. 495-501 ◽  
Author(s):  
Steven J. Damon ◽  
Russell L. Groves ◽  
Michael J. Havey

Natural variation exists in onion (Allium cepa L.) for amounts of epicuticular waxes on foliage, and plants with lower amounts of these waxes suffer less feeding damage from onion thrips (Thrips tabaci Lind.). Wild-type onion possesses copious amounts of epicuticular waxes and is often referred to as “waxy.” The recessively inherited “glossy” phenotype has significantly less wax relative to waxy types and shows resistance to onion thrips but is vulnerable to spray damage, foliar pathogens, and excessive transpiration. Phenotypes visually intermediate between waxy and glossy also exist in onion, which we refer to as “semiglossy.” Epicuticular waxes on the leaves of glossy, semiglossy, and waxy onions were evaluated for appearance using scanning electron microscopy (SEM) and amounts and types were analyzed using gas chromatography/mass spectrometry. Wax crystals were clearly visible on the surface of waxy foliage with decreasing amounts on semiglossy and none on glossy leaves. The ketone hentriacontanone-16 was the most prevalent wax on leaves of waxy onion and was significantly (P < 0.01) less on semiglossy relative to waxy plants and on glossy relative to waxy and semiglossy plants. Numbers of adult and immature onion thrips were significantly reduced (P < 0.05) on glossy and/or semiglossy accessions relative to waxy in field and greenhouse cage experiments. These results indicate that semiglossy plants possess intermediate amounts of epicuticular waxes that may protect leaves from diseases or environmental stresses while still conferring resistance to onion thrips. Therefore, the semiglossy phenotype should be useful in integrated programs managing this important onion pest.


Author(s):  
Michael J. Havey ◽  
Derek J. Hunsaker ◽  
Eduardo D. Munaiz

The amounts and types of epicuticular waxes on onion (Allium cepa) leaves affect the severity of feeding damage by onion thrips (Thrips tabaci), a serious insect pest of onion. Onion plants with light green leaves are referred to as “glossy” and accumulate less epicuticular wax relative to the blue–green (“waxy”) foliage of wild-type onion. The onion cultivar Odourless Greenleaf (OGL) has visually glossy foliage, shows resistance to thrips feeding damage, and has the unique profile of accumulating waxes with 28 or fewer carbons. Plants of glossy OGL were crossed with the glossy inbred B9885 and waxy inbred lines DH2107, DH066619, and B8667. Hybrid progenies from glossy OGL by waxy plants had waxy foliage, indicating recessiveness of the glossy OGL phenotype relative to the waxy phenotype. Hybrids from the cross of glossy OGL with glossy B9885 were also waxy, revealing different genetic bases for the glossy phenotype in these two onions. Hybrid plants were self-pollinated and segregations in F2 families from OGL × waxy crosses fit the expected 3:1 ratio for the single locus at which the homozygous recessive genotype conditions glossy foliage. Segregations in F2 families from crosses of glossy 9885 × glossy OGL fit the 9:7 ratio, supporting two independently segregating loci, where the recessive genotype at either locus conditions the glossy phenotype. Amounts and types of epicuticular waxes on leaves of F2 progenies from crosses of OGL × waxy B8667 and glossy B9885 × OGL were determined using gas chromatography-mass spectrometry. Single-nucleotide polymorphisms were genotyped and genetic maps were constructed. The visually glossy phenotype from OGL and its unique profile of epicuticular waxes were conditioned by one locus on chromosome 6, for which we propose the name glogl. Onion populations such as OGL with unique epicuticular wax profiles will be important germplasms for the development of onion cultivars that suffer less feeding damage from onion thrips compared with waxy onion.


2014 ◽  
Vol 139 (5) ◽  
pp. 597-602 ◽  
Author(s):  
Steven J. Damon ◽  
Michael J. Havey

Natural variation exists in onion (Allium cepa L.) for amounts and types of epicuticular waxes on leaves. Wild-type waxy onion possesses copious amounts of these waxes, whereas the foliage of semiglossy and glossy phenotypes accumulates significantly less wax. Reduced amounts of epicuticular waxes have been associated with resistance to onion thrips (Thrips tabaci Lindeman), an important insect pest of onion. A segregating family from the cross of waxy and semiglossy onions was used to map single nucleotide polymorphisms (SNPs) and identify chromosome regions affecting amounts and types of epicuticular waxes as measured by gas chromatography–mass spectrometry. The amount of the primary epicuticular wax on onion leaves, hentriacontanone-16, was controlled by one region on chromosome 5. One region on chromosome 2 affected concentrations of several primary fatty alcohols. Results indicate that the region on chromosome 2 may be associated with the acyl reduction pathway, and the region on chromosome 5 may affect the decarbonylation pathway of epicuticular wax biosynthesis. Because lower amounts of epicuticular waxes are recessively inherited, SNPs tagging regions on chromosomes 2 and 5 will be useful for marker-assisted breeding to vary amounts and types of epicuticular waxes on onion foliage with the goal to develop cultivars resistant to onion thrips.


2020 ◽  
Vol 145 (1) ◽  
pp. 30-35 ◽  
Author(s):  
Eduardo D. Munaiz ◽  
Russell L. Groves ◽  
Michael J. Havey

Onion thrips (Thrips tabaci) is the main insect pest of onion (Allium cepa), and feeding damage routinely causes serious yield losses. Lower amounts of epicuticular waxes on onion leaves have been associated with fewer onion thrips and less feeding damage, and research is needed to assess the relationships between amounts and composition of epicuticular waxes and feeding damage by onion thrips. This study used gas chromatography mass spectroscopy to determine amounts and types of epicuticular waxes on the foliage of onion accessions that had been field-selected for lower damage from onion thrips. Hentriacontanone-16 (H16), octaconasol-1, and triacontanol-1 were the most prevalent waxes on the foliage of these selections. Amounts of H16 were significantly lower on selections visually classified as having glossy or semiglossy foliage. Semiglossy selections were identified with similar amounts of total epicuticular wax as waxy phenotypes, due primarily to lower amounts of H16 and higher amounts of other waxes. These semiglossy selections suffered significantly less feeding damage from onion thrips in a field evaluation, supporting the identification of unique wax profiles toward the development of thrips-resistant onion.


2020 ◽  
Vol 145 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Eduardo D. Munaiz ◽  
Michael J. Havey

Onion (Allium cepa) plants with lower amounts of epicuticular waxes on foliage suffer less damage from the insect pest Thrips tabaci (onion thrips). Glossy onion accumulates significantly less epicuticular wax compared with wild-type “waxy” onion, and a single recessive locus (gl) has been proposed to condition this phenotype. Genetic analyses of types and amounts of epicuticular waxes were completed using two segregating families from the cross of the glossy inbreds B9885 and B9897 (both originally selected from the onion cultivar White Persian) with waxy inbred B8667 and semiglossy (intermediate amounts of waxes) inbred B5351, respectively. F2 progenies were grown in greenhouses and scored visually for foliar phenotypes, and amounts and types of epicuticular waxes were determined using gas chromatography-mass spectrometry (GCMS). For one F2 family from the cross of glossy B9885 by waxy B8667, visually scored glossy vs. waxy foliage fit a 1:3 ratio and the phenotype mapped to chromosome 8 of onion. This same region on chromosome 8 was significantly associated with amounts of the ketone hentriacontanone-16 (H16) and fatty alcohols 1-octacosanol (Oct1) and 1-triacontanol (Tri1). Visually scored F2 progeny from the cross of glossy B9897 × semiglossy B5351 did not fit expected models for one or two recessive loci. Significant quantitative trait loci (QTL) were revealed on chromosomes 5 and 8 controlling amounts of H16. Epistasis was detected between regions on chromosomes 1 and 8, and a 100-fold increase of H16 was conditioned by homozygous genotypes for the B5351 region on chromosome 1 and the B9885 region on chromosome 8. The three QTL model explained 41% of the phenotypic variation for amounts of H16 at logarithm of odds of 16.6. Amounts of Oct1 and Tri1 in the B9897 × B5351 family were associated with a major QTL on chromosome 1, explaining 37% to 46% of the phenotypic variation, respectively. This research demonstrates that glossy foliage of ‘White Persian’ onion is conditioned by a recessive locus on chromosome 8 for which we propose the name glwp. These results are important for selection of onion with unique profiles of epicuticular waxes to reduce losses resulting from onion thrips.


1991 ◽  
Vol 37 (4) ◽  
pp. 540-546 ◽  
Author(s):  
Linda Thienpont ◽  
Lothar Siekmann ◽  
Alexander Lawson ◽  
Elisabeth Colinet ◽  
Andrȳ De Leenheer

Abstract The Community Bureau of Reference of the European Communities has produced four batches of lyophilized serum Certified Reference Materials, two for cortisol (CRM 192 and 193) and two for progesterone (CRM 347 and 348). For cortisol, one of the pools consisted of serum from healthy blood donors, whereas the second batch was supplemented with pure cortisol. The progesterone Reference Materials contained only endogenous hormone concentrations. Assessment of vial-to-vial variability in the cortisol and progesterone concentrations showed no between-sample inhomogeneity, and the materials were stable. The quality of the materials was therefore considered sufficient for certification of the values for the cortisol and progesterone concentrations by a collaborative study involving several laboratories from the European Communities, using isotope dilution gas chromatography-mass spectrometry. Inaccuracy in reconstitution of the lyophilized materials was less than 0.3%; imprecision of sampling was less than 0.2%. For determinations of cortisol and progesterone concentrations, the mean within-laboratory coefficients of variation (CVs) were 1.76% (CRM 192), 1.19% (CRM 193), 1.64% (CRM 347), and 1.75% (CRM 348). The between-laboratory CVs were greater: CRM 192, 1.79%; CRM 193, 1.48%; CRM 347, 2.08%; and CRM 348, 2.16%. The concentrations in the reconstituted Reference Materials were certified to be 273 nmol/L in CRM 192 and 763 nmol/L in CRM 193 for cortisol and 10.13 nmol/L in CRM 347 and 40.3 nmol/L in CRM 348 for progesterone. Uncertainties at the 0.95 confidence level--6 (CRM 192), 14 (CRM 193), 0.21 (CRM 347), and 1.0 nmol/L (CRM 348)--were considered compatible with the intended use of the materials.


1997 ◽  
Vol 122 (6) ◽  
pp. 788-791 ◽  
Author(s):  
Joyce G. Latimer ◽  
Ray F. Severson

Epicuticular waxes were analyzed to explain the visible differences in the waxy bloom of conditioned broccoli (Brassica oleracea L. Group Italica `Green Duke') transplants. Seedlings (22 days old) were subjected to brushing (40 cycles per minute, 1 minute twice daily), wind (7 m·s-1 for 5 minutes twice daily), or moisture-stress conditioning (MSC; visible wilt for 2 to 4 hours daily) for 16 (1987) or 21 (1988) days during transplant production in the greenhouse. The epicuticular waxes of the uppermost fully expanded leaves were removed by dipping detached leaves into methylene chloride. The extract was derivatized with trimethylsilyl reagents and subjected to capillary gas chromatography. The primary epicuticular wax components were the nonpolar C29 compounds nonacosane, nonacosan-15-ol, and nonacosan-15-one, which were identified by mass spectrometry. In a Summer 1987 experiment, cuticle samples taken over time of treatment indicated acclimation to the conditioning treatments relative to untreated plants. After 9 days of treatment, the amount of total epicuticular waxes present on the leaves was reduced 38%, 31%, or 11% by wind, brushing, or MSC, respectively. However, after 15 days of treatment, the amount of cuticle present was reduced 15% by brushing but only 6% by wind and was 17% greater in MSC-treated plants. Two weeks after transplanting to the field there were no differences in the amount or composition of the epicuticular waxes. In Fall 1988, all treatments reduced plant growth, but only MSC tended to increase the amount of C29 epicuticular components during greenhouse production. Differences in the amounts of epicuticular waxes were no longer significant after 8 days in the field.


1994 ◽  
Vol 77 (4) ◽  
pp. 917-924 ◽  
Author(s):  
Roger T Wilson ◽  
Joseph M Groneck ◽  
Kathleen P Holland ◽  
A Carolyn Henry

Abstract A gas chromatographic/mass spectrometric procedure is described for the quantitation and confirmation of clenbuterol residues from cattle, sheep, and swine tissues. After liquid–liquid extraction and derivatization with phosgene in an aqueous pH 10.1 buffer, the cyclic oxazolidone derivative is quantitated with a clenbuterol analogue as internal standard (NAB-760 CI). Confirmation is accomplished by comparison of ion ratios with those of a pure synthesized standard of clenbuterol oxazolidin-3-one obtained by selected ion monitoring, electron ionization gas chromatography/mass spectrometry on a benchtop instrument. Statistical information based on a series of standard curves for fortified tissues is included to describe method performance. Ion ratio variations were under 15%, and coefficients of variation for spiked tissue standard curves were above 0.997. Recoveries averaged 87.1 ± 6.6% for liver tissues across all 3 species and 67.1 ± 3.8% for muscle tissue across all 3 species.


Molecules ◽  
2020 ◽  
Vol 25 (15) ◽  
pp. 3454 ◽  
Author(s):  
Eduardo D. Munaiz ◽  
Philip A. Townsend ◽  
Michael J. Havey

Epicuticular waxes on the surface of plant leaves are important for the tolerance to abiotic stresses and plant–parasite interactions. In the onion (Allium cepa L.), the variation for the amounts and types of epicuticular waxes is significantly associated with less feeding damage by the insect Thrips tabaci (thrips). Epicuticular wax profiles are measured using used gas chromatography mass spectrometry (GCMS), which is a labor intensive and relatively expensive approach. Biochemical spectroscopy is a non-destructive tool for measurement and analysis of physiological and chemical features of plants. This study used GCMS and full-range biochemical spectroscopy to characterize epicuticular waxes on seven onion accessions with visually glossy (low wax), semi-glossy (intermediate wax), or waxy (copious wax) foliage, as well as a segregating family from the cross of glossy and waxy onions. In agreement with previous studies, GCMS revealed that the three main waxes on the leaves of a wild type waxy onion were the ketone hentriacontanone-16 (H16) and fatty alcohols octacosanol-1 (Oct) and triacontanol-1 (Tri). The glossy cultivar “Odourless Greenleaf” had a unique phenotype with essentially no H16 and Tri and higher amounts of Oct and the fatty alcohol hexacosanol-1 (Hex). Hyperspectral reflectance profiles were measured on leaves of the onion accessions and segregating family, and partial least-squares regression (PLSR) was utilized to generate a spectral coefficient for every wavelength and prediction models for the amounts of the three major wax components. PLSR predictions were robust with independent validation coefficients of determination at 0.72, 0.70, and 0.42 for H16, Oct, and Tri, respectively. The predicted amounts of H16, Oct, and Tri are the result of an additive effect of multiple spectral features of different intensities. The variation of reflectance for H16, Oct, and Tri revealed unique spectral features at 2259 nm, 645 nm, and 730 nm, respectively. Reflectance spectroscopy successfully revealed a major quantitative trait locus (QTL) for amounts of H16, Oct, and Tri in the segregating family, agreeing with previous genetic studies. This study demonstrates that hyperspectral signatures can be used for non-destructive measurement of major waxes on onion leaves as a basis for rapid plant assessment in support of developing thrips-resistant onions.


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