Field application of vesicular-arbuscular mycorrhizal fungi improved garlic yield in disinfected soil

 The production of certified garlic propagation material requires measures to be taken against pathogenic nematodes. Methyl bromide (MB) may be used for this purpose, but is known to cause stunting in Allium spp. Vesicular-arbuscular mycorrhizal (VAM) fungal inoculum was applied to the planting furrow after MB treatment. VAM-inoculated plants were larger, had more green leaves, an increased photosynthesis rate, especially at low light intensities, and higher fresh and dry weights than plants in uninoculated plots. The mean bulb weights from uninoculated and VAM-treated plots were 27 g and 51 g respectively. The native or an improved VAM population should be reintroduced after soil disinfection to ensure satisfactory garlic yields. Accepted: 15 January 1997     

Transformation and regeneration of garlic (Allium sativum L.) by Agrobacterium-mediated gene transfer

 By using highly regenerative calluses, we developed a stable transformation system in garlic (Allium sativum L.). The temperature and number of days of co-cultivation with Agrobacterium tumefaciens was shown to be an important factor in transient expression of the uid A gene. After a culture period of 5 months in selection medium containing hygromycin, 20 shoots were induced from ca. 1000 calluses, among which 15 plants expressed β-glucuronidase activity upon staining with X-Gluc. Shoots developed into transgenic garlic after 1 month. Integration of the uid A gene was confirmed by Southern blot analysis for genomic DNA of transgenic garlic plants. Received: 25 October 1999 / Revision received: 16 February 2000 / Accepted: 22 February 2000     

Regeneration of garlic callus as affected by clonal variation, plant growth regulators and culture conditions over time

 A long-term regeneration system for garlic (Allium sativum L.) clones of diverse origin was developed. Callus was initiated on a modified Gamborg's B-5 medium supplemented with 4.5 μM 2,4-D and maintained on the same basal medium with 4.7 μM picloram+0.49 μM 2iP. Regeneration potential of callus after 5, 12 and 16 months on maintenance medium was measured using several plant growth regulator treatments. The 1.4 μM picloram+13.3 μM BA treatment stimulated the highest rate of shoot production. Regeneration rate decreased as callus age increased, but healthy plantlets from callus cultures up to 16-months-old were produced for all clones. Regeneration of long-term garlic callus cultures could be useful for clonal propagation and transformation. Received: 24 September 1998 / Revision received: 27 January 1999 / Accepted: 26 February 1999     

Unlocking variability: inherent variation and developmental traits of garlic plants originated from sexual reproduction

Recent collections of fertile garlic (Allium sativum) accessions from Central Asia allow a detailed study of seedling developments and the evaluation of inherent variations. We hereby provide a comprehensive account of the ontogenesis of a population of garlic seedlings and their vegetative and reproductive traits. A nucleotide binding site profiling marker technology was applied to provide conclusive evidence for the cross-pollination nature of garlic, and to compare the levels of polymorphism between progeny derived from a single mother clone fertilized by several pollinators. The seedlings’ population demonstrates a large variation in vegetative and reproductive characters, including bulbing ability, bulb color and size, clove number, and response to environmental conditions, similar to that of the genepool of vegetatively propagated garlic clones. In addition, a large variation in flowering and seed production ability was recorded. The understanding of garlic physiology, the availability of the large variability unleashed by sexual reproduction, and the possible utilization of sexual hybridization opens the way for genetic studies and breeding work. Haim D. Rabinowitch and Rina Kamenetsky contributed equally to this paper.     

Mitochondrial and total DNA RAPD patterns can distinguish restorers of CMS lines in sorghum

 Seven sorghum restorer lines that differentially restore (or maintain) the A1 and A2 cytoplasmic male-sterile (CMS) cytoplasms were studied by RFLP analyses of their mtDNAs and RAPD analyses of their mitochondrial DNA (mtDNA) and total DNA to understand nuclear mitochondrial combinations that are present in these lines. RFLP data from 11 mitochondrial gene probes were inadequate to classify these seven lines. However, the analysis of RAPD profiles of total DNA could distinguish these lines on the basis of their ability to restore completely or partially the fertility in the A1/A2 CMS cytoplasms. Interestingly, RAPD profiles of mtDNAs of these lines also followed the same pattern as that of the total DNA. These results indicate that the different restorer lines possess specific nuclear-cytoplasm combinations. Further, the results also show that the RAPD technique can be used to identify markers for different cytoplasms used in CMS. Received: 26 August 1997 / Accepted: 9 October 1997     

Identification and characterisation of resistance against rust (Puccinia allii) in garlic (Allium sp.) germplasm

In this work we studied the resistance response against Puccinia allii of 533 Allium sp. germplasm accessions under field conditions. Ten resistant accessions were selected to further characterise the defence mechanisms operative. Histological studies showed a range of defence mechanisms, acting alone or combined, that impeded fungal development at different stages. Some accessions showed prehaustorial resistance. In other accessions, mesophyll cells were penetrated by the fungus, but then hypersensitive response leading to cell death hampered fungal development. In some cases, cell death was very fast and early colony abortion was obvious already by 2 days after inoculation (DAI), whereas in others it was slow being marked only by 6 DAI. The fact that resistant accessions studied showed both pre‐ and posthaustorial resistance offers breeding opportunities for these traits.     

Molecular mapping of the rf1 gene for pollen fertility restoration in sorghum (Sorghum bicolor L.)

We report the molecular mapping of a gene for pollen fertility in A1 (milo) type cytoplasm of sorghum using AFLP and SSR marker analysis. DNA from an F₂ population comprised of 84 individuals was screened with AFLP genetic markers to detect polymorphic DNAs linked to fertility restoration. Fifteen AFLP markers were linked to fertility restoration from the initial screening with 49 unique AFLP primer combinations (+3/+3 selective bases). As many of these AFLP markers had been previously mapped to a high-density genetic map of sorghum, the target gene (rf1) could be mapped to linkage group H. Confirmation of the map location of rf1 was obtained by demonstrating that additional linkage group-H markers (SSR, STS, AFLP) were linked to fertility restoration. The closest marker, AFLP Xtxa2582, mapped within 2.4 cM of the target loci while two SSRs, Xtxp18 and Xtxp250, flanked the rf1 locus at 12 cM and 10.8 cM, respectively. The availability of molecular markers will facilitate the selection of pollen fertility restoration in sorghum inbred-line development and provide the foundation for map-based gene isolation. Received: 22 August 2000 / Accepted: 18 October 2000     

Insecticidal activity of garlic juice in two dipteran pests

Abstract

• 1 Botanical products excluding pyrethroids constitute a small, but growing portion of the U.K. pesticides market. With increasing legislative pressure upon chemical pesticides such as organophosphates, interest in this sector is increasing steadily.
• 2 Garlic Allium sativum L. juices and extracts form the basis of several commercially available pest control products, but the performance of these products is variable, possibly due to lack of quality control upon batches of materials in the manufacturing process.
• 3 Some garlic products designed for use in the food industry are subjected to rigorous batch‐control to ensure organoleptic consistency. We studied the insecticidal efficacy of a commercially produced food grade garlic juice using two target dipteran pests, Delia radicum (L.) and Musca domestica L.
• 4 Exposure of the two species to different concentrations of garlic juice revealed variability in insecticidal effect across life stages. LC₅₀ values recorded for D. radicum were: eggs (7‐day exposure) 0.8%; larvae (24‐h exposure) 26.4%; larvae (48‐h exposure) 6.8%; and adults (24‐h exposure) 0.4%. LC₅₀ values recorded for M. domestica were: eggs (7‐day exposure) 1.6%; larvae (24‐h exposure) 10.1%; larvae (24‐h exposure) 4.5%; and adults (24‐h exposure) 2.2%.
• 5 Mortality rates caused by the garlic juice were comparable with those obtained with the organophosphate pesticide Birlane^®, indicating parity of effect at various concentrations depending on life stage.
• 6 We conclude that this product may provide an effective, naturally‐derived insecticide for use in agricultural systems against dipteran pests.

     

高产SOD大蒜悬浮细胞系的选育

筛选获得了高产 SOD大蒜悬浮细胞系 ,培养 1 8d后达到最大生物量及最大 SOD总酶活分别为2 2 .1 2 g.DW/ L及 1 0 .81× 1 0 4 U/ L,最大单位细胞酶活为 679.67U/ g.FW,SOD最大比活力可达 85 .98U/mg Pro,具有较强的 SOD合成能力。     

A PCR-marker for the CMS(1) inducing cytoplasm in chives derived from recombination events affecting the mitochondrial gene atp9

The complete coding and 3′-flanking region of the mitochondrial gene atp9 of chives (Allium schoenoprasum L.) was determined in order to develop primers that allow the identification of atp9-related sequences in subsequent PCR-amplifications. One of these sequences is of a chimerical nature, consisting of atp9-homologous regions on its end, interrupted by an insertion that is composed of one atp6-homologous part and one part of unknown origin. This PCR-fragment is 762 bp in size and exclusively amplified in the sterility inducing cytoplasm of CMS₁. Thus it can be used as a PCR-marker in order to distinguish this cytoplasm type from the remaining cytoplasm types of chives. The chimerical marker sequence forms a putative open reading frame (orfA501), from which CMS₁ might originate. Received: 6 June 2001 / Accepted: 3 August 2001     

Genetic variability in callus formation and regeneration of garlic (Allium sativum L.)

Twenty garlic Allium sativum (L.) genotypes were analysed for genetic variation in their ability to form callus (in one medium) and regenerate shoots (in four different media). Genotypes showed significant differences in the analysis of variance of all the traits tested. The accession Printanor showed the best general behaviour, with 83% callus-producing explants, 44.7% organogenic explants, and 15.35 shoots/g of callus. The best regeneration medium was MBO, without growth regulators. Shoot production capacity was examined with the additive main effects and multiplicative interaction model that proved to be a powerful tool for analysis and easy comprehension of the strong genotype×medium interactions frequently observed in in vitro culture systems. Received: 5 February 1998 / Revision received: 11 May 1998 / Accepted: 1 June 1998     

Non-random mating in classical lekking grouse species: seasonal and diurnal trends

This paper is the first to integrate both field and theoretical approaches to demonstrate that fertility benefits can be a direct benefit to females mating on the classical lek. Field data collected for male sharp-tailed grouse (Tympanuchus phasianellus), a classical lekking species, revealed potential fertility benefits for selective females. Adult males and individuals occupying centrally located territories on the lek were found to have significantly larger testes than juveniles and peripheral individuals. Further, using empirical data from previously published studies of classical lekking grouse species, time-series analysis was employed to illustrate that female mating patterns, seasonal and daily, were non-random. We are the first to show that these patterns coincide with times when male fertility is at its peak. Received: 26 February 1999 / Revised: 13 December 1999 / Accepted: 15 March 2000     

Detection of somaclonal variation in garlic (Allium sativum L.) using RAPD and cytological analysis

Plants were regenerated by somatic embryogenesis from long-term callus cultures derived from five garlic (Allium sativum L.) cultivars. Thirty-five of these plants were subjected to RAPD analysis. The frequency of variation was found to be cultivar dependent: approximately 1% in the two clones Solent White and California Late and around 0.35% in another three clones, Chinese, Long Keeper and Madena. Certain band changes were found in regenerants of different cultivars, suggesting the existence of a mutation-sensitive part of the garlic genome. The karyotypes of another 75 regenerants derived from the same callus cultures of three parental garlic clones were examined. Of these plants, 9.3% were found to be tetraploids, 4% aneuploid and 2.6% showed a change in the position of the secondary constriction. No association could be shown between the rate of variation for molecular and cytological characters either by comparing cultivars or examining individual regenerants. Received: 30 July 1996 / Revision received: 28 October 1996 / Accepted: 12 November 1996     

Diversity among male-sterility-inducing and male-fertile cytoplasms of onion

Hybrid-onion (Allium cepa) seed is produced using systems of cytoplasmic-genic male sterility (CMS). Two different sources of CMS (S and T cytoplasms) have been genetically characterized. Testcrosses of N-cytoplasmic maintaining and restoring genotypes to S and T cytoplasmic lines demonstrated that different alleles, or loci, restore male fertility for these two male-sterile cytoplasms. Other sources of CMS have been used or reported in Europe, Japan and India, and their relationships to S and T cytoplasms are not clear. Restriction fragment length polymorphisms were identified in the organellar genomes among commercially used male-sterile cytoplasms from Holland, Japan and India, and were compared to S and T cytoplasms. Mitochondrial DNA diversity among 58 non-S-cytoplasmic open-pollinated onion populations was also assessed. All five putative CMS lines selected from the Indian population Nasik White Globe were identical to S cytoplasm for all polymorphisms in the chloroplast genome, and always possessed the same-sized mitochondrial fragments as S cytoplasm. T cytoplasm, the male-sterile cytoplasm used to produce the Dutch hybrid Hygro F₁, and two sources of CMS from Japan, were similar and showed numbers of mitochondrial polymorphisms similar to those observed among the 58 non-S-cytoplasmic open-pollinated populations. This research demonstrates that the same, or very similar, male-sterile cytoplasms have been independently isolated and exploited for hybrid-seed production in onion. Received: 27 October 1999 / Accepted: 12 February 2000     

A low-density genetic map of onion reveals a role for tandem duplication in the evolution of an extremely large diploid genome

 The bulb onion, Allium cepa L., is a diploid (2n=2x=16) plant with a huge nuclear genome. Previous genetic and cytogenetic analyses have not supported a polyploid origin for onion. We developed a low-density genetic map of morphological markers, randomly amplified polymorphic DNAs (RAPD), and restriction fragment length polymorphisms (RFLP) as a tool for onion improvement and to study the genome organization of onion. A mapping population of 58 F₃ families was produced from a single F₁ plant from the cross of two partially inbred lines (Brigham Yellow Globe 15-23 and Alisa Craig 43). Segregations were established for restoration of male fertility in sterile cytoplasm, complementary light-red bulb color, 14 RAPDs, 110 RFLPs revealed by 90 anonymous cDNA clones, and 2 RFLPs revealed by a cDNA clone of alliinase, the enzyme responsible for the characteristic Allium flavors. Duplicated RFLP loci were detected by 21% of the clones, of which 53% were unlinked (>30 cM), 5% loosely linked (10–30 cM), and 42% tightly linked (<10 cM). This duplication frequency is less than that reported for paleopolyploids but higher than for diploid species. We observed 40% dominant RFLPs, the highest yet reported among plants. Among duplicated RFLP loci, 19% segregated as two loci each with two codominant alleles, 52% segregated as one locus with codominant alleles and one locus with only a dominant fragment, and 29% segregated as two loci with only dominant fragments. We sequenced cDNAs detecting duplicated RFLPs; 63% showed homology to known gene families (e.g., chlorophyll binding proteins, ubiquitin, or RuBISCO), and 37% were unique clones showing significant homology to known genes of low-copy number or no homology to database sequences. Duplicated RFLPs showing linkage could be due to retroviral-like sequences in adjacent coding regions or intrachromosomal, as opposed to whole genome, duplications. Previous cytological analyses and this genetic map support intrachromosomal duplication as a mechanism contributing to the huge onion genome. Received: 3 July 1997 / Accepted: 8 August 1997     

High frequency shoot regeneration and plantlet formation from root tip of garlic

An efficient and novel method of direct shoot regeneration from root tips in garlic was developed. The influence of growth regulators, basal media and age of root explant on shoot initiation and proliferation was examined. The best growth regulator combination was 1-naphthaleneacetic acid and 6-benzyladenine at 1 and 10 μM, respectively, inducing shoot initiation from 75% of the explants. The frequency of shoot initiation on different basal media was similar. Explant root tips from plantlets taken 15 to 18 days after sprouting showed the highest shoot initiation (95%). In contrast to Murashige and Skoog medium, which produced more than 10 shoots per explant, B5 medium produced smaller shoots, although the number was higher. Rooting of individual shoots was induced after transfer to medium without growth regulators. Plantlets, after acclimatization in a growth cabinet, were successfully transplanted to the field, and no phenotypic variation was observed among them. The technique has potential applicability for rapid propagation of garlic. This revised version was published online in June 2006 with corrections to the Cover Date.     

Intercropping garlic at different planting times and densities for insect pest or crop yield and value management in tobacco fields

Although the densities of tobacco pests have been decreased in garlic‐tobacco fields, further studies are needed to judge the effects of garlic transplanting densities or times on tobacco pests in tobacco fields. Therefore, field experiments were conducted in Liancheng County in Longyan City, Fujian Province, in China in 2014 and 2015. Myzus persicae (Sulzer) abundance, the species or abundance ratios of enemies and pests, the intercropping effects and the tobacco yield and crop value showed that the effects of transplanting tobacco 10 days after garlic transplantation at a density of 5.85 individual plants per square meter on pests were stronger than those of other treatments. Aphid abundance was significantly lower in transplanting tobacco 10 days after garlic transplantation at a density of 5.85 individual plants per square meter than in the other treatments. The ratio between enemies and pests in transplanting tobacco 10 days after garlic transplantation at a density of 5.85 individual plants per square meter was higher than those in the other treatments. The intercropping effects of transplanting tobacco 10 days after garlic transplantation at a density of 5.85 individual plants per square meter on Myzus persicae, Spodoptera litura (Fabricius), Heliothis assulta Guenee and Nezara viridula Linnaeus were significantly stronger than those of the other treatments, whereas the effects of transplanting tobacco 15 days after garlic transplantation at a garlic density of 5.85 individual plants per square meter on Agrotis ypsilon (Rottemberg) were significantly stronger. Additionally, the yield and crop value of transplanting tobacco 10 days after garlic was transplanted at a density of 5.85 individual plants m^?2 were higher than those of the other treatments. Therefore, our study demonstrated that the model of transplanting tobacco 10 days after garlic was transplanted at a density of 5.85 individual plants m^?2 is an optimal management strategy to control flue‐cured tobacco pests and to acquire higher crop yield in garlic‐tobacco fields.