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1.
G. Suzuki M. Watanabe T. Nishio 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2000,101(1-2):80-85
In Brassica, self-incompatibility genes SLG (for S-locus glycoprotein) and SRK (for S-receptor kinase) are located in the S-locus complex region with several other S-linked genes. The S locus is a highly polymorphic region: polymorphism has been observed not only in sequences of SLG and SRK but also in the location of the S-locus genes. In order to compare the physical location of the S-locus genes in various S haplotypes, we used six class-I S haplotypes of B. rapa and seven class-I S haplotypes of B. oleracea in this study. DNA gel blot analysis using pulsed-field gel electrophoresis (PFGE) showed that the physical distances between
SLG and SRK in B. rapa are significantly shorter than those in B. oleracea and that the sizes of MluI and BssHII fragments harboring SLG and SRK are less variable within B. rapa than within B. oleracea. We concluded that several large genomic fragments might have been inserted into the S-locus region of B. oleracea after allelic differentiation of S-locus genes.
Received: 20 September 1999 / Accepted: 8 October 1999 相似文献
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Yoshinobu Takada Takayuki Nakanowatari Jun Sato Katsunori Hatakeyama Tomohiro Kakizaki Akiko Ito Go Suzuki Hiroshi Shiba Seiji Takayama Akira Isogai Masao Watanabe 《Sexual plant reproduction》2005,17(5):211-217
Plants have evolved many systems to prevent inappropriate fertilization. Among them, incompatibility is a well-organized system in which pollen germination or pollen-tube growth is inhibited in pistils. Self-incompatibility (SI), rejecting self-pollen, promotes outbreeding in flowering plants. On the other hand, inter-species incompatibility, preventing gene flow among species to restrict outbreeding, usually occurs unilaterally, and is known as unilateral incompatibility (UI). In Brassicaceae, little is known about the molecular mechanism of UI, although S-locus genes involved in recognition of self-pollen have been characterized in the SI system. In the present study, we characterized novel UI observed between members of the same species, Brassica rapa; pollen of Turkish SI lines was specifically rejected by pistils of the Japanese commercial SI variety Osome. The incompatible phenotype of this intra-species UI closely resembled that of SI. Segregation analysis revealed that the pollen factor of this UI was not linked to the S-locus.The revised version was published online in December 2004 with corrections to figure 1. 相似文献
5.
M. Iwano K. Sakamoto G. Suzuki M. Watanabe S. Takayama K. Fukui K. Hinata A. Isogai 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,96(6-7):751-757
The physical localization of the S-glycoprotein (SLG) locus in the chromosome of Brassica campestris L. ‘pekinensis’ cv ‘Kukai’ was visualized by multi-color fluorescent in situ hybridization (McFISH). ‘Kukai’, which is an F1 hybrid between two parental lines, T-17 and T-18, has two SLG genes from both T-17 and T-18. In this study, a 1.3-kb DNA fragment was amplified from the genomic DNA of T-17 by PCR using
a set of primers specific to the class-I SLG. From the genomic DNA of T-18, no DNA fragment was amplified using these primers. In the genomic Southern hybridization,
a cloned PCR product hybridized with the genomic DNA of T-17 or F1 but not with that of T-18. The PCR product had a sequence homology of approximately, 85% to another class-I SLG gene, SLG-9. Therefore, the PCR product from T-17 was named SLG-17, as it is thought to be a member of the class-I SLG. Using SLG-17 as the probe, FISH was carried out to visualize the position of the SLG locus. McFISH was also carried out simultaneously using the SLG-17 and SLG-9 genes as probes. The SLG-17 gene was detected as a doublet signal at the interstitial region close to the end of a small chromosome, with the signal
site being identical to that of SLG-9. Therefore, it is concluded that the SLG-17 gene is localized at the interstitial region close to the end of the chromosome derived from T-17 in Brassica campestris L. ‘pekinensis’ cv ‘Kukai’.
Received: 18 September 1997 / Accepted: 6 October 1997 相似文献
6.
Snowdon RJ Friedrich T Friedt W Köhler W 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(4):533-538
Oilseed rape (Brassica napus L.) is an amphidiploid species that originated from a spontaneous hybridisation of Brassica rapa L. (syn. campestris) and Brassica oleracea L., and contains the complete diploid chromosome sets of both parental genomes. The metaphase chromosomes of the highly homoeologous
A genome of B. rapa and the C genome of B. oleracea cannot be reliably distinguished in B. napus because of their morphological similarity. Fluorescence in situ hybridisation (FISH) with 5S and 25S ribosomal DNA probes
to prometaphase chromosomes, in combination with DAPI staining, allows more dependable identification of Brassica chromosomes. By comparing rDNA hybridisation and DAPI staining patterns from B. rapa and B. oleracea prometaphase chromosomes with those from B. napus, we were able to identify the putative homologues of B. napus chromosomes in the diploid chromosome sets of B. rapa and B. oleracea, respectively. In some cases, differences were observed between the rDNA hybridisation patterns of chromosomes in the diploid
species and their putative homologue in B. napus, indicating locus losses or alterations in rDNA copy number. The ability to reliably identify A and C genome chromosomes
in B. napus is discussed with respect to evolutionary and breeding aspects.
Received: 13 July 2001 / Accepted: 23 August 2001 相似文献
7.
A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and
the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected
with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic
plants of all three cultivars used were obtained with frequencies of 1.6–2.7% when the explants were regenerated in shoot
regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that
transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion.
Received: 15 December 1998 / Revision received: 2 July 1999 · Accepted: 8 July 1999 相似文献
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Gene transfer between canola (Brassica napus L. and B. campestris L.) and related weed species 总被引:1,自引:0,他引:1
Brassica species are particularly receptive to gene transformation techniques. There now exists canola genotypes with transgenic herbicide resistance for glyphosate, imidazolinone, sulfonylurea and glufosinate herbicides. The main concern of introducing such herbicide resistance into commercial agriculture is the introgression of the engineered gene to related weed species. The potential of gene transfer between canola (Brassica napus and B. campestris) and related weed species was determined by hand pollination under controlled greenhouse conditions. Canola was used as both male and female parent in crosses to the related weed species collected in the Inland Northwest region of the United States. Weed species used included: field mustard (B. rapa), wild mustard (S. arvensis) and black mustard (B. nigra). Biological and cytological aspects necessary for successful hybrid seed production were investigated including: pollen germination on the stigma; pollen tube growth down the style; attraction of pollen tubes to the ovule; ovule fertilisation; embryo and endosperm developmental stages. Pollen germination was observed in all 25 hybrid combinations. Pollen tubes were found in the ovary of over 80% of combinations. About 30% of the hybrid combinations developed to the heart stage of embryo development or further. In an additional study involving transgenic glufosinate herbicide resistant B. napus and field mustard it was found that hybrids occurred with relatively high frequency, hybrids exhibited glufosinate herbicide resistance and a small proportion of hybrids produced self fertile seeds. These fertile plants were found to backcross to either canola or weed parent. 相似文献
10.
B. R. Choudhary P. Joshi 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,102(6-7):1123-1128
The crossability between Brassica tournefortii (TT, 2n = 20) and Brassica rapa (AA, 2n = 20) and the cytomorphology of their F1 hybrids were studied. Hybrids between these two species were obtained only when B. tournefortii was involved as a female parent. The hybrid plants were intermediate for most of the morphological attributes and were found
to be free from white rust under field conditions. The F1 plants showed poor pollen fertility, although occasional seed set was achieved from open pollination. Self-pollination or
backcrosses did not yield any seeds in these plants. The occurrence of chromosome association ranging from bivalents (0–7),
trivalents (0–2) to a rare quadrivalent (0–1) in the dihaploid hybrids indicates pairing between the T and A genomes. The
homoeologous pairing coupled with seed set in the F1 plants offer an opportunity for interspecific gene transfers from B. tournefortii to B. rapa and vice-versa through interspecific hybridization.
Received: 3 July 2000 / Accepted: 22 September 2000 相似文献
11.
TTG1(Transparent Testa Glabra 1)蛋白是一种WD40类蛋白,参与植物的生长和发育。采用RT-PCR方法从芜菁品种‘津田'中克隆了BrTTG1 cDNA序列(GenBank登录号HM208590)。该基因cDNA开放阅读框长度为1 014 bp,编码一个由337个氨基酸残基组成的蛋白,该蛋白分子量为37.28 kDa,理论等电点为4.66。与其他植物中的TTG1蛋白进行同源性比对结果显示,BrTTG1与甘蓝型油菜的TTG1同源性最高。BrTTG1蛋白在31~337位氨基酸处含有WD40超家族的保守结构域。荧光定量PCR检测BrTTG1在‘津田'芜菁不同组织中的表达结果表明,该基因在有花青素合成的红色‘津田'芜菁根皮中表达量最高。 相似文献
12.
Greenhouse-grown plants of turnip rape Brassica rapa ssp. oleifera (syn. B. campestris) cv. Valtti and Sisu were transformed by Agrobacterium tumefaciens infection. Of the three A. tumefaciens strains tested (C58C1, EHA105 and LBA4404), LBA4404 gave the best results. Segments excised from one to two upper internodes
of an inflorescence-carrying stem served as explants for the Agrobacterium infection. Cultivation of the explants horizontally during the first 3 days of co-cultivation with A. tumefaciens following immediate selection of transformed tissue of the stem segments placed vertically basal side down were critical.
Use of silver nitrate (5–10 mg/l) in the culture medium and Micropore (3 M) paper tape for sealing plates was also beneficial.
Transgenic shoots were recovered using either hygromycin or kanamycin (20–25 mg/l) selection. Hygromycin was preferable, as
the proportion of `escapes' was 90% under kanamycin and 10% under hygromycin selection. Regeneration was achieved by culturing
the explants for 3–6 days on 0.5 mg/l of 2,4-di-chlorophenoxyacetic acid and 1–2 weeks on 2–3 mg/l of 6-benzyl aminopurine
with/without 0.05 mg/l α-naphthaleneacetic acid. Recovered shoots were then cultured on hormone-free MS medium. This culture program gave 60–80% shoot
regeneration. Regenerants were tested by histological β-glucuronidase staining and Southern blotting. The recovery rate of transgenic shoots was 4–9% of the number of explants used
in the experiments.
Received: 28 November 1997 / Revision received: 25 March 1998 / Accepted: 22 November 1998 相似文献
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Improved resistance to bacterial soft rot by protoplast fusion between Brassica rapa and B. oleracea
J. P. Ren M. H. Dickson E. D. Earle 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2000,100(5):810-819
Erwinia soft rot is a destructive disease of Brassica rapa vegetables. Reliable sources of resistance and control methods are limited, so development of highly resistant breeding lines
is desirable. Protoplasts from B. rapa and B. oleracea genotypes selected for resistance to soft rot were fused in order to combine different sources of resistance. Twelve somatic
hybrids (synthetic B. napus) were obtained and confirmed by morphology, nuclear DNA content, and RAPD analysis. They were normal looking plants that
easily set seeds following self-pollination and backcrossing to B. rapa. Assays of detached leaves or seedlings inoculated in a mist-chamber showed that most somatic hybrids had lower disease severity
ratings than the B. rapa fusion partner and a commercial variety of B. napus. Some progeny from selfing or backcrossing of somatic hybrids to B. rapa showed much more resistance than either fusion partner. The offspring populations of the somatic hybrids (F1–S1 and F1–BC1) clearly moved to the resistant direction compared to the parents; the percentage of resistant plants increased from 21%
(average of parents) to 36% (F1–S1) and 48% (F1–BC1). These results suggest that it may be possible to obtain highly resistant B. rapa lines by further backcrossing and selection.
Received: June 1999 / Accepted: 29 July 1999 相似文献
15.
K. Srinivasan V. G. Malathi P. B. Kirti S. Prakash V. L. Chopra 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(5-6):976-981
Monosomic chromosome addition lines of Brassica oxyrrhina in the background of alloplasmic B. campestris carrying B. oxyrrhina cytoplasm were generated and characterised through morphology, cytology and molecular (RAPD) analysis. Four successive backcrosses
of the synthetic alloploid B. oxycamp with B. campestris yielded 24 monosomic addition plants that were grouped into seven different synteny groups based on morphological similarity
and RAPD patterns. Each synteny group exhibited morphological features diagnostic for the presence of individual B. oxyrrhina chromosomes including some novel phenotypes. Meiotic studies of the addition lines revealed the homoeology of four B. oxyrrhina chromosomes (synteny groups 1, 3, 5 and 6 ) with B. campestris chromosomes as indicated by trivalent associations, with the highest homoeology (44.23%) in synteny group 1 and the lowest
(6.1%) in synteny group 3. Seed fertility of the addition lines ranged from 94.85% (synteny group 1) to 56.98% (synteny group
5). All of the addition lines were male-sterile except synteny group 6 which had 12–16% stainable pollen. Ovule transmission
of the B. oxyrrhina chromosomes added to the progenies of addition lines ranged from 23.52% (synteny group 6) to 14% (synteny group 7). RAPD
analysis confirmed the validity of synteny grouping based on morphological observations. Approximately 45% of the primers
studied were informative, giving B. oxyrrhina-specific RAPD bands unique for each synteny group, except group 6.
Received: 20 October 1997 / Accepted: 31 March 1998 相似文献
16.
An effective plant regeneration procedure and a gene transfer system via Agrobacterium tumefaciens were developed in Brassica campestris ssp. parachinensis. Hypocotyls from 5-day-old seedlings with 2 days pre-culture were infected with Agrobacterium strain MOG301 harboring a binary vector containing a synthetic Bacillus thuringiensis (B.t.) cry1Ab or cry1Ac gene with full codon-modification. After culture and selection on MS medium supplemented with 4.0 mg/l BAP, 2.0 mg/l NAA,
70 μM AgNO3 and 50 mg/l kanamycin, a number of kanamycin-resistant plantlets were regenerated. PCR and Southern blotting analysis were
used to identify and characterize the transgenic plants with the integrated cry1Ab or cry1Ac gene. Western blotting analysis of the transgenic plants confirmed the expression of insecticidal proteins encoded by cry1Ab or cry1Ac. Subsequent bioassay with larvae of the Diamondback moth, Plutella xylostella, demonstrated that the transgenic plants were resistant to feeding damage.
Received: 22 February 1999 / Revision received: 14 April 1999 / Accepted: 26 April 1999 相似文献
17.
Expression of GFP and Bt transgenes in Brassica napus and hybridization with Brassica rapa 总被引:4,自引:0,他引:4
M. D. Halfhill H. A. Richards S. A. Mabon C. N. Stewart 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,103(5):659-667
It is possible to monitor the movement of transgenes by tagging them with green fluorescent protein (GFP). In order to develop
a model to study transgene flow, canola (Brassica napus cv Westar) was transformed with two GFP constructs, mGFP5er (GFP only) and pSAM 12 [GFP linked to a synthetic Bacillus thuringiensis (Bt) cry1Ac endotoxin gene]. Transformed callus sectors that fluoresced green were preferentially selected in the tissue culture process.
Four independent GFP canola events and 12 events of GFP/Bt canola were regenerated through tissue culture. GFP fluorescence
was macroscopically detectable throughout the entire life cycle of canola. The GFP/Bt events were insecticidal to neonate
corn earworm (Helicoverpa zea) larvae and prevented herbivory damage. Fluorescence intensity at 508 nm varied between the independent transformation events,
and ranged from 7.6×105 to 13.8×105 (counts per second) in contrast with the wild-type at 5.3×105 cps. Nine GFP/Bt and three GFP events were hybridized with three wild accessions of B. rapa. The resultant hybrids fluoresced green and were insecticidal to neonate corn earworm larvae to the same degree as the transgenic
canola parents. However, fluorescence intensities of the hemizygous F1 hybrid lines were lower than the respective original homozygous canola parents. Each F1 hybrid line was backcrossed by hand onto the B. rapa parent, and transgenic backcrosses were produced at rates ranging from 15% to 34%. These data suggest that GFP can be used
as a tool to monitor transgene flow from crop species to wild relatives.
Received: 11 September 2000 / Accepted: 1 February 2001 相似文献
18.
K. Song M. K. Slocum T. C. Osborn 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1995,90(1):1-10
Construction of a detailed RFLP linkage map of B. rapa (syn. campestris) made it possible, for the first time, to study individual genes controlling quantitative traits in this species. Ninety-five F2 individuals from a cross of Chinese cabbage cv Michihili by Spring broccoli were analyzed for segregation at 220 RFLP loci and for variation in leaf, stem, and flowering characteristics. The number, location, and magnitude of genes underlying 28 traits were determined by using an interval mapping method. Zero to five putative quantitative trait loci (QTL) were detected for each of the traits examined. There were unequal gene effects on the expression of many traits, and the inheritance patterns of traits ranged from those controlled by a single major gene plus minor genes to those controlled by polygenes with small and similar effects. The effect of marker locus density on detection of QTL was analyzed, and the results showed that the number of QTL detected did not change when the number of marker loci used for QTL mapping was decreased from 220 to 126; however, a further reduction from 126 to 56 caused more than 15% loss of the total QTL detected. The detection of putative minor QTL by removing the masking effects of major QTL was explored. 相似文献
19.
M. Beschorner V. Heberer W. Odenbach 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(5):885-890
Thirty resynthesized Brassica napus lines with defined S-allele constitution and the ancestral B. oleracea and B. campestris lines were used for the analysis of S- locus glycoproteins (SLGs). The aim of this study was to investigate (1) whether the
S-specific glycoproteins of the diploid ancestor lines were also expressed in the amphidiploid hybrids and (2) whether the
occurrence of SLG bands was correlated with the activity of the respective S-alleles, which had been tested by means of diallele
pollination tests in a previous study. Stigma proteins were separated by isoelectric focusing (IEF)-gel electrophoresis, and
glycoprotein bands were identified by Western blotting and Con-A/peroxidase reaction. The SLG bands of the B. campestris parent could be detected in all 30 resynthesized B. napus lines. In contrast, B. oleracea SLG bands could only be detected in 12 resynthesized B. napus lines. Only B. napus lines which carried the dominant B. oleracea S-alleles S8 and S29 showed respective SLG bands in all cases. Nine B. napus lines showed only glycoprotein bands of the B. campestris parent, although the biological functioning of the B. oleracea S-alleles was demonstrated by test-pollinations. New SLG bands different from those of the B. oleracea and B. campestris parents occurred in 16 B. napus lines. The expression level of the SLGs in B. napus was not correlated with the self-incompatibility phenotype, not only in the case of recessive S-alleles (S2, S15), but also for dominant alleles (e.g. S14, S32, S45).
Received: 22 January 1999 / Accepted: 30 January 1999 相似文献
20.
W. Y. Cheung G. Champagne N. Hubert L. Tulsieram D. Charne J. Patel B. S. Landry 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(1-2):73-82
Self-incompatibility (SI) in Brassica is a sporophytic system, genetically determined by alleles at the S-locus, which prevents self-fertilization and encourages outbreeding. This system occurs naturally in diploid Brassica species but is introduced into amphidiploid Brassica species by interspecific breeding, so that in both cases there is a potential for yield increase due to heterosis and the
combination of desirable characteristics from both parental lines. Using a polymerase chain reaction (PCR) based analysis
specific for the alleles of the SLG (S-locus glycoprotein gene) located on the S-locus, we genetically mapped the S-locus of B. oleracea for SI using a F2 population from a cross between a rapid-cycling B. oleracea line (CrGC-85) and a cabbage line (86-16-5). The linkage map contained both RFLP (restriction fragment length polymorphism)
and RAPD (random amplified polymorphic DNA) markers. Similarly, the S-loci were mapped in B. napus using two different crosses (91-SN-5263×87-DHS-002; 90-DHW-1855-4×87-DHS-002) where the common male parent was self-compatible,
while the S-alleles introgressed in the two different SI female parents had not been characterized. The linkage group with the S-locus in B. oleracea showed remarkable homology to the corresponding linkage group in B. napus except that in the latter there was an additional locus present, which might have been introgressed from B. rapa. The S-allele in the rapid-cycling Brassica was identified as the S29 allele, the S-allele of the cabbage was the S
5 allele. These same alleles were present in our two B. napus SI lines, but there was evidence that it might not be the active or major SI allele that caused self-incompatibility in these
two B. napus crosses.
Received: 7 June 1996/Accepted: 6 September 1996 相似文献