Genetic variation of petaloid male-sterile cytoplasm of carrots revealed by sequence-tagged sites (STSs)

The mitochondrial DNA of various carrot lines was characterized by random amplified polymorphic DNA (RAPD) analysis, and six sequence-tagged sites (STSs) led to identification of the petaloid type of cytoplasmic male sterility (CMS). Using six STS primer combinations, we were able to classify five CMS lines into two groups and eight fertile carrots into six groups. Both the STS1 and the STS4 primer combinations differentiated CMS cytoplasms from the fertile cytoplasms, and the STS2 primer combination revealed two different types of CMS cytoplasms – of Wisconsin Wild and Cornell origins. Cybrid carrot lines with petaloid flowers which had been obtained by asymmetric cell fusion could also be separated from fertile cybrids by the STS1 primer combination. The STS1 fragment contained a homologous sequence with the orfB gene. DNA gel blot analysis indicated that homologous regions to the STS1 fragment existed in fertile types as well as the CMS types, although the restriction fragment size patterns differed. These observations demonstrate that rearrangements involving this region occurred in the mitochondrial genome. The STS4 fragment had a more complicated gene structure, including retrotransposon-like sequences and small segments of chloroplast genome. Received: 10 September 1998 / Accepted: 24 February 1999     

Using physical barriers to prevent carrot fly (Psila rosae (Fabricius)) damage in domestic production

A field experiment was used to assess the effectiveness of different barriers in protecting garden‐scale carrot production from carrot fly (Psila rosae (Fabricius)) damage. Some of the vertical barriers tested were found to provide a useful method of protecting early season carrots from carrot fly in terms of the percentage of carrots free from damage but, under cumulative pest pressure of several generations of carrot fly, such barriers were found to provide insufficient protection. Gardeners should therefore completely cover their carrot crop to attain an acceptable level of control, and this was found to be especially important for carrots harvested later in the season. There were positive effects of some barrier types on yield which may be due, at least in part, to the protection given by the barriers to carrot seedlings.     

Variation in sequence and RNA editing within core domains of mitochondrial group II introns among plants

The 3' regions of several group II introns within the mitochondrial genes nad1 and nad7 show unexpected sequence divergence among flowering plants, and the core domains 5 and 6 are predicted to have weaker helical structure than those in self-splicing group II introns. To assess whether RNA editing improves helical stability by the conversion of A-C mispairs to A-U pairs, we sequenced RT-PCR amplification products derived from excised intron RNAs or partially spliced precursors. Only in some cases was editing observed to strengthen the predicted helices. Moreover, the editing status within nad1 intron 1 and nad7 intron 4 was seen to differ among plant species, so that homologous intron sequences shared lower similarity at the RNA level than at the DNA level. Plant-specific variation was also seen in the length of the linker joining domains 5 and 6 of nad7 intron 3; it ranged from 4 nt in wheat to 11 nt in soybean, in contrast to the 2-4 nt length typical of classical group II introns. However, this intron is excised as a lariat structure with a domain 6 branchpoint adenosine. Our observations suggest that the core structures and sequences of these plant mitochondrial introns are subject to less stringent evolutionary constraints than conventional group II introns.     

Relationship between the concentration of chlorogenic acid in carrot roots and the incidence of carrot fly larval damage

Chlorogenic acid (1·24-3·36 mg/g) was identified as the main phenolic component in the peel of carrots by hplc analysis. The higher the concentration of chlorogenic acid in different cultivars the greater the susceptibility to carrot fly larval damage. Increases in concentration were found both after carrot fly damage and after carrots had overwintered in the field. The presence and location of chlorogenic acid was confirmed in sections of carrot tissues, mounted in 0·05 M ammonia solution by viewing them using a u.v.-epifluorescent microscope. The importance of phenolic compounds and their function in the production of insect cuticle is discussed in relation to the different concentrations of chlorogenic acid and resistance to carrot fly in carrots.     

Bionomics of the carrot fly Psila rosae F.; the infestation and sampling of carrot crops

The headlands of a carrot field are generally more heavily attacked by carrot fly larvae than the remainder of the field.
The methods of sampling a carrot crop are outlined: it has been found that, for fields up to 10 acres, samples of carrots taken in alternate twos and threes in the midfield, and in twos on the headlands, give a reliable measure of the infestation. For larger fields the number of carrots is doubled. Transects of headlands and whole fields have been made, and are useful in showing the actual amount of damage in a crop. Thus it is possible to represent each field as a three-dimensional body showing the amount and distribution of the attack.
The deterioration of attacked carrot crops during autumn and winter has been followed. Deterioration is measured by (1) percentage carrots attacked, (2) number of mines per 100 carrots and (3) the percentage of carrots unsaleable or unfit for market. The relations between (1) mines per 100 carrots and time, (2) percentage attack and time, (3) mines per 100 carrots and percentage attack, and (4) mines per 100'carrots and percentage unsaleability are stated. Based on the above relationships, methods of prediction of deterioration have been worked out.
The importance of shelter in determining the degree and position of attack is discussed and the relative effects of different types of shelter on infestation are compared.     

Divergent intron conservation in the mitochondrial nad2 gene: signatures for the three bryophyte classes (mosses,liverworts, and hornworts) and the lycophytes

The slow-evolving mitochondrial DNAs of plants have potentially conserved information on the phylogenetic branching of the earliest land plants. We present the nad2 gene structures in hornworts and liverworts and in the presumptive earliest-branching vascular land plant clade, the Lycopodiopsida. Taken together with the recently obtained nad2 data for mosses, each class of bryophytes presents another pattern of angiosperm-type introns conserved in nad2: intron nad2i1 in mosses; intron nad2i3 in liverworts; and both introns, nad2i3 and nad2i4, in hornworts. The lycopods Isoetes and Lycopodium show diverging intron conservation and feature a unique novel intron, termed nad2i3b. Hence, mitochondrial introns in general are positionally stable in the bryophytes and provide significant intraclade phylogenetic information, but the nad2 introns, in particular, cannot resolve the interclade relationships of the bryophyte classes and to the tracheophytes. The necessity for RNA editing to reconstitute conserved codon entities in nad2 is obvious for all clades except the marchantiid liverworts. Finally, we find that particularly small group II introns appear as a general feature of the Isoetes chondriome. Plant mitochondrial peculiarities such as RNA editing frequency, U-to-C type of RNA editing, and small group II introns appear to be genus-specific rather than gene-specific features.     

An eco-metabolomic study of host plant resistance to Western flower thrips in cultivated,biofortified and wild carrots

Domestication of plants and selection for agronomic traits may reduce plant secondary defence metabolites relative to their ancestors. Carrot (Daucus carota L.) is an economically important vegetable. Recently, carrot was developed as a functional food with additional health-promoting functions. Biofortified carrots contain increased concentrations of chlorogenic acid as an antioxidant. Chlorogenic acid is involved in host plant resistance to Western Flower Thrips (Frankliniella occidentalis), one of the key agri- and horticultural pests worldwide. The objective of this study was to investigate quantitative host plant resistance to thrips in carrot and to identify candidate compounds for constitutive resistance. As such we explored whether cultivated carrot is more vulnerable to herbivore attack compared to wild carrot. We subjected a set of 14 biofortified, cultivated and wild carrot genotypes to thrips infestation. We compared morphological traits and leaf metabolic profiles of the three most resistant and susceptible carrots using nuclear magnetic resonance spectroscopy (NMR). In contrast to our expectation, wild carrots were not more resistant to thrips than cultivated ones. The most thrips resistant carrot was the cultivar Ingot which is known to be tolerant against carrot root fly (Psila rosae). Biofortified carrots were not resistant to thrips. Plant size, leaf area and number of leaf hairs did not differ between resistant and susceptible carrots. The metabolic profiles of the leaves of resistant carrots were significantly different from those of susceptible carrots. The leaves of resistant carrots contained higher amounts of the flavanoid luteolin, the phenylpropanoid sinapic acid and the amino acid β-alanine. The negative effect of these compounds on thrips was confirmed using in-vitro bioassays. Our results have potential implications for carrot breeders. The natural variation of metabolites present in cultivated carrots can be used for improvement of thrips resistance. This is especially promising in view of the candidate compounds we identified since they do not only confer a negative effect on thrips but as antioxidants also play an important role in the improvement of human health.     

Further studies relating chlorogenic acid concentration in carrots to carrot fly damage

The effects of first generation carrot fly larval damage on chlorogenic acid concentration in carrots was investigated in a field experiment at Wellesbourne in 1985. In a separate experiment carrots grown in the absence of a resident population of carrot fly were also analysed for chlorogenic acid; these carrots maintained low concentrations of chlorogenic acid through the summer and autumn until low ground temperatures occurred from November to January. The relationship between chlorogenic acid concentration and damage by the first generation of carrot fly was described by a similar model to the one derived previously for late-generation damage but without the cultivar dependence. This may have been because first generation damage takes place in mid-summer when soil temperature is not sufficiently low for differential chlorogenic acid production by carrot cultivars. The model supports the hypothesis that carrot fly damage increases chlorogenic acid production which subsequently encourages further attack. The increase in acid production due to the low winter temperature may be the mechanism which, in turn, induced a differential cultivar response in carrots harvested during the winter.     

Introgression and persistence of cultivar alleles in wild carrot (Daucus carota) populations in the United States

Premise

Cultivated species and their wild relatives often hybridize in the wild, and the hybrids can survive and reproduce in some environments. However, it is unclear whether cultivar alleles are permanently incorporated into the wild genomes or whether they are purged by natural selection. This question is key to accurately assessing the risk of escape and spread of cultivar genes into wild populations.

Methods

We used genomic data and population genomic methods to study hybridization and introgression between cultivated and wild carrot (Daucus carota) in the United States. We used single nucleotide polymorphisms (SNPs) obtained via genotyping by sequencing for 450 wild individuals from 29 wild georeferenced populations in seven states and 144 cultivars from the United States, Europe, and Asia.

Results

Cultivated and wild carrot formed two genetically differentiated groups, and evidence of crop–wild admixture was detected in several but not all wild carrot populations in the United States. Two regions were identified where cultivar alleles were present in wild carrots: California and Nantucket Island (Massachusetts). Surprisingly, there was no evidence of introgression in some populations with a long-known history of sympatry with the crop, suggesting that post-hybridization barriers might prevent introgression in some areas.

Conclusions

Our results provide support for the introgression and long-term persistence of cultivar alleles in wild carrots populations. We thus anticipate that the release of genetically engineered (GE) cultivars would lead to the introduction and spread of GE alleles in wild carrot populations.