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1.
Kui Shin Voo Clayton L. Rugh Joe C. Kamalay 《In vitro cellular & developmental biology. Plant》1991,27(3):117-124
Summary We describe a tissue culture procedure for somatic embryogenesis and plantlet regeneration in cotton (Gossypium hirsutum L. cv. Coker 312). Callused explants or individual globular embryos were transferred to basal media to induce somatic embryogenesis.
To determine characteristic early indicators of successful germination and conversion, we identified six types of embryos
that developed on basal media. Two of the six embryo types, designated as tulip-shaped and trumpet-shaped, could undergo conversion
in preliminary tests, whereas the others had little or no developmental potential. Several media treatments designed to enhance
the maturation of globular somatic embryos failed to increase the fraction of embryos which matured to form recoverable types.
In efforts to improve plantlet recovery, tulip-shaped embryos were used in limited trials to contrast the effects of chemical
and physical desiccation treatments on germination and conversion. The selective use of tulip-shaped somatic embryos, coupled
with partial desiccation, seems to have augmented plant recovery. Growth habit, flowering, seed set, and lint production of
most of the regenerated plants were comparable to seed-derived plants grown under the same conditions.
Partial research support was provided by state and federal funds appropriated to the Ohio Agricultural Research and Development
Center, The Ohio State University. 相似文献
2.
Transformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants 总被引:2,自引:0,他引:2
Ebrahim Firoozabady David L. DeBoer Donald J. Merlo Edward L. Halk Lorraine N. Amerson Kay E. Rashka Elizabeth E. Murray 《Plant molecular biology》1987,10(2):105-116
Cotton (Gossypium hirsutum L.) cotyledon tissues have been efficiently transformed and plants have been regenerated. Cotyledon pieces from 12-day-old aseptically germinated seedlings were inoculated with Agrobacterium tumefaciens strains containing avirulent Ti (tumor-inducing) plasmids with a chimeric gene encoding kanamycin resistance. After three days cocultivation, the cotyledon pieces were placed on a callus initiation medium containing kanamycin for selection. High frequencies of transformed kanamycin-resistant calli were produced, more than 80% of which were induced to form somatic embryos. Somatic embryos were germinated, and plants were regenerated and transferred to soil. Transformation was confirmed by opine production, kanamycin resistance, immunoassay, and DNA blot hybridization. This process for producing transgenic cotton plants facilitates transfer of genes of economic importance to cotton. 相似文献
3.
L. S. Kaushik D. P. Singh R. S. Paroda 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1984,68(6):487-491
Summary The data from an experiment in cotton consisting of three testers and 12 lines selected deliberately have been analysed. The investigation showed higher specific combining ability variance for yield of seed cotton and number of bolls, indicating the predominance of non-additive gene action. Of parental lines, H777 was found to possess high g.c.a. effects for seed cotton yield, number of bolls and number of sympodes. Parent H842 contributed only for boll weight, whereas H655 was good general combiner for number of monopodes. There appeared to be better chances for increasing the yield by exploiting hybrid vigour for the number of bolls and boll weight. The presence of marked non-additive gene effects, in addition to additive gene effects, indicated the need for exploiting both the fixable and non-fixable components of genetic variance for increasing productivity in cotton. 相似文献
4.
Tissue culture methods for improvement of cotton has lagged seriously compared to other major crops. A method for regeneration of cotton which includes a morphogenetically competent cell suspension was needed to facilitate selection of stress-resistant variants and gene manipulation. Preliminary screening of eight strains of Gossypium hirsutum L. for embryogenic potential resulted in the production of somatic embryos in all strains. Coker 312 was selected for use in the development of a model regeneration system for G. hirsutum. Calli were initiated from hypocotyl tissues of 3-day-old-seedlings. Globular embryos were present after six weeks in culture. Calli were subcultured to liquid suspension in growth regulator-free medium. After three to four weeks, suspensions were sieved to collect globular and heart stage embryos. Collected embryos developed further when plated onto semi-solid medium. To induce germination and plantlet growth, mature embryos were placed on sterile vermiculite saturated with medium. Upon development of roots and two true leaves, plantlets were potted in peat and sand, and hardened. Mature plants and progeny have been obtained with this procedure. A high percentage of infertile plants was observed among the regenerants.Abbreviations NAA
1 naphthaleneacetic acid
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- MS
Murashige and Skoog
- BA
6 benzylamino purine
- 2i P
N6-(2-isopentenyladenine 相似文献
5.
A. P. Tyagi 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,74(2):280-283
Summary Two diverse parents of upland cotton namely J.34 and I.C. 1926 were crossed. A comparison between biparental intermated progenies and F3 families indicated alteration of correlation coefficient between yield and halo length. The significant negative correlation in F3 population between these two attributes changed to a positive but non significant one in biparental intermated progenies. A change in correlation coefficients was expected due to breakage of linkage upon intermating. An increase in the correlation coefficients could also be expected when linkages are predominantly in the repulsion phase. It is suggested that intermating in early generations coupled with selection of desirable segregants may prove a useful method for improving yield and quality simultaneously. The diallel selective mating system may also supplement intermating to improve yield and quality in cotton.Part of Ph.D. Thesis submitted to the Haryana Agricultural University. Hissar-125004, India 相似文献
6.
In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.) 总被引:5,自引:0,他引:5
D. C. Agrawal A. K. Banerjee R. R. Kolala A. B. Dhage A. V. Kulkarni S. M. Nalawade S. Hazra K. V. Krishnamurthy 《Plant cell reports》1997,16(9):647-652
Induction of multiple shoots in cotton (Gossypium hirsutum L. cv. Anjali-LRK 516) has been achieved with cotyledonary nodes devoid of cotyledons and apical meristems. Explants from 35-day-old seedlings yielded the maximum number of shoots (4.7 shoots/explant) using Murashige and Skoog (MS) basal medium supplemented with 6-benzylaminopurine and kinetin (2.5 mg/1 each). Explants from 35-day-old seedlings raised in glass bottles produced a higher number of multiple shoots (8.3 shoots/explant) than those grown in glass tubes and cultured on the same shoot induction medium. Elongation of multiple shoots was obtained on liquid or agar MS basal medium without phytohormones. In vitro shoots were rooted on half-strength agar-solidified MS basal medium or with 0.05 or 0.1 mg/1 naphthaleneacetic acid. Hardening and survival of tissue culture plantlets was 95% under greenhouse conditions.Abbreviations
BAP
6-Benzylaminopurine
- GA3
Gibberellic acid
- MS
Murashige and Skoog medium
-
NAA
-Napthaleneacetic acid 相似文献
7.
Summary The growth of submerged cotton (Gossypium hirsutum L.) fibers from cultured ovules has been investigated. The results indicate that exogenous plant hormone levels regulate
the induction of submerged fiber growth. The age of ovules at induction is also important. Cell diameter, wall thickness,
and cell length of submerged fibers were measured and compared with air-grown fibers and fibers grown in vivo (produced by cotton plants grown in the greenhouse). Various cellwall thickening patterns were observed among submerged fibers,
while only one predominant cell-wall deposition pattern was produced in air-grown fibers and in fibers produced in vivo. The diameter of submerged fibers was about the same as that of air-grown fibers but about 22% less than that of fibers grown,
in vivo. It appears that the secondary cell wall thickenings are initiated earlier in submerged fibers. The cell-wall thickness of
submerged fibers, at 41 d post anthesis (DPA), was 51% greater than that of fibers grown in vivo, whereas the cell-wall thickness of air-grown fibers was 42% less than that of fibers produced in vivo. The cell length of submerged fibers was approximately half that of fibers grown in vivo. and the air-grown fiber length was about two-thirds of fibers grown in vivo. The age of ovules at induction affects the outcome of the air-grown fiber-cell length, but does not appear to affect the
length of submerged fiber cells. To produce submerged fiber growth, we found that the optimal age of ovules at induction was
0 DPA, and the optimal medium (with a GA3 of 0.5 μM and an IAA range of 5-20 μM) depends on the time of ovule induction (−2 to+2DPA). We conclude that conditions leading to submerged cotton fiber growth
have great potential for (a) direct monitoring of growth and making precise, detailed measurements during fiber growth and
development; (b) producing cellulose and fibers in vitro more efficiently than earlier ovule-culture methods; and (c) using these unique cultures to obtain a better understanding
of signal transduction and gene expression leading to growth, development, and programmed cell death in the life history of
the cotton fiber. 相似文献
8.
R. G. Dani R. J. Kohel 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,77(4):569-575
Summary The nature of gene action and of maternal influence governing cottonseed oil attributes were determined with four lines, two each with high and low seed-oil percentage. For this purpose, P1, P2, F0, F1, F2 and alternative sets of BC1 and BC2 generations were analysed in six cross-combinations and their reciprocals. Marginal extents of heterosis for seed-oil percentage were noticeable in F1, with inbreeding depression in F2. Data from reciprocal backcrosses provided evidence in favour of maternal rather than cytoplasmic effects of seed-oil development. Relatively higher extents of heterosis, sizeable inbreeding depression and reciprocally unequal F2 averages were characteristic of the seed index trait, which often showed a reversal of effects from F1 to F2. Reverse reciprocal backcrosses exhibited some differences, including greater resemblance between the types, (A/B)A and (B/A)A, in addition to variable dose effects in seed index. Thus, the differences between F1 seed index values were not due to cytoplasmic influence. Positive heterotic effects for seed-oil index, especially among the backcrosses, ranged between 16.08% and 47.29% over midparent averages. Genetic component estimates from analysis of similar sets of crosses differing only in reciprocal backcrosses, and also from sets of reciprocal crosses between any two parental combinations, were inconsistent. Scaling tests detected presence of epistasis within and between a majority of cross-combinations. Despite reciprocal differences, additive gene effects for seed-oil percentage were significant in 7 out of 24 crosses, representing high x low, low x high and low x low seed-oil parents. Those were, however, accompanied by significant dominance effects of higher order. In crosses involving low seed-oil percentage parents SA1060 and SA229, all six components were detected significant, with opposite effects of dominance and dominance x dominance epistatic components. Significant additive components were also detected for seed index and seed-oil index in 7 and 5 out of 24 crosses, respectively. In the inheritance of seed index and seed-oil index, dominance effects were more important. Epistatic components of additive x additive, and to a lesser extent, those of dominant x dominant were found significant. 相似文献
9.
Tanoh Hilaire Kouakou Pierre Waffo Téguo Josep Valls Yatty Justin Kouadio Alain Decendit Jean-Michel Mérillon 《Plant Cell, Tissue and Organ Culture》2006,86(3):405-409
For the first time, trans-resveratrol, a stilbene, has been identified in cotton cell suspensions. Cell suspensions of Coker 312, a cultivar which produces embryogenic structures, acccumulate trans-resveratrol contrary to those of cultivar R405-2000, which do not. This stilbene may be a good phenolic marker for induction of somatic embryogenesis in cotton. 相似文献
10.
Plant regeneration via somatic embryogenesis in many cultivars of cotton (Gossypium hirsutum L.) 总被引:4,自引:0,他引:4
Summary Embryogenic callus was formed from several cultivars of cotton (Gossypium hirsutum L.) when sections of hypocotyl and cotyledon were cultured on medium supplemented with 5 mg/liter 6-(γ, γ-dimethylallyl-amino)-purine
(2iP) and 0.1 mg/liter α-naphthaleneacetic acid (NAA) for callus initiation and proliferation, and subcultured on medium supplemented
with 5 mg/liter NAA and 0.1 to 1 mg/liter 2iP for embryogenic callus induction. It seems that a high 2iP:auxin ratio is preferred
for callus initiation and proliferation, but should be exchanged with a higher NAA:cytokinin ratio before differentiation
will occur. Embryogenic calluses were recovered at a frequency of 2 to 85% depending on the cultivar used. Coker cultivars
produced embryogenic callus faster and at higher frequencies than other cultivars. Embryogenic callus produced somatic embryos
on phytohormone-free medium. This medium was used to maintain and proliferate embryogenic callus for a perid of 18 to 24 mo.
Somatic embryos were converted to plants on a lower ionic strength medium supplemented with 0.1 mg/liter gibberellic acid
(GA3) and 0.01 mg/liter NAA. Glucose was the only carbohydrate used through all phases of tissue culture and was much better than
sucrose, on which phenolic production was very high. High temperature (30° C) and low light intensity (9 μE · m−2 · s−1) were optimal conditions for callus initiation, embryogenic callus induction, and maintenance, whereas lower temperature
(25° C) and high light intensity (90 μE · m−2 s−1) were the optimal conditions for somatic embryo maturation, germination, and plantlet development. Plants could be regenerated
within 10 to 12 wk in Cokers or 7 to 8 mo. in others. 相似文献
11.
Summary The storage protein content of somatic embryos of Gossypium hirsutum L. cv. Coker 201 was determined using extinction level, antigen/antibody association detection methods. Mature storage protein was first detected in early globular-stage somatic embryos at a total concentration of 0.36% of the embryo protein mass. Tulip-stage and mature somatic embryos were comprised of 3.0% and 1.3% mature storage protein, respectively. Maximum storage protein synthesis was found to occur during early globular- and early heart-stages. During this period of development, significant levels of protein precursors were found also to accumulate. The pattern of storage protein synthesis, processing and accumulation paralleled the pattern that has been reported for the zygotic system, although somatic embryos accumulate storage protein at much earlier stages and to a lesser degree. The possibility of using complex biochemical pathways to monitor embryogenic systems in vitro is discussed. 相似文献
12.
An efficient grafting system for transgenic plant recovery in cotton (Gossypium
hirsutum L.) 总被引:1,自引:0,他引:1
Shuangxia Jin Shaoguang Liang Xianlong Zhang Yichun Nie Xiaoping Guo 《Plant Cell, Tissue and Organ Culture》2006,85(2):181-185
A successful transformation program relies on the number of survival plants in soil that can be obtained. Low recovery of transgenic plants is still a key restrictive factor for transgenic cotton production. In order to utilize genetic transformation in cotton breeding program effectively, an efficient grafting system for recovering plants derived from somatic embryogenesis following Agrobacterium infection and kanamycin selection was developed. Various aspects of in vitro grafting were examined in efforts to improve the efficiency of transformant recovery. Using strong seedling rootstocks was the first important step to obtain high rate of successful grafts. Scion size >0.6 cm and seedling rootstock at age of 6–12 days were appropriate for grafting. The successful grafting ratio was higher when using hypocotyls without radicle. Shoot-tip and shoot stem with axillary bud were also suitable for in vitro grafting, which meant we could significantly improve the survival ratio of transgenic plantlets, because one plantlet has a shoot-tip but several axillary buds. Based on our data, the period from in vitro seedling rootstock germination to transplant of grafts to field usually takes one month. Over 90% successful grafting ratio could be obtained under optimal conditions, which represented a significant improvement over currently available methods for recovery of cotton plantlet from somatic embryogenesis after transformation. Ex vitro grafting could also be used for plant recovery, which gave an average of successful grafting ratio of 71.9%. However, this method was strongly affected by environmental factors. 相似文献
13.
Construction of a comprehensive PCR-based marker linkage map and QTL mapping for fiber quality traits in upland cotton (Gossypium hirsutum L.) 总被引:1,自引:0,他引:1
Zheng-Sheng Zhang Mei-Chun Hu Jian Zhang Da-Jun Liu Jing Zheng Ke Zhang Wei Wang Qun Wan 《Molecular breeding : new strategies in plant improvement》2009,24(1):49-61
To facilitate marker assisted selection, there is an urgent need to construct a saturated genetic map of upland cotton (Gossypium hirsutum L.). Four types of markers including SSR, SRAP, morphological marker, and intron targeted intron–exon splice junction (IT-ISJ)
marker were used to construct a linkage map with 270 F2:7 recombinant inbred lines derived from an upland cotton cross (T586 × Yumian 1). A total of 7,508 SSR, 740 IT-ISJ and 384
SRAP primer pairs/combinations were used to screen for polymorphism between the two mapping parents, and the average polymorphisms
of three types of molecular markers represented 6.8, 6.6 and 7.0%, respectively. The polymorphic primer pairs/combinations
and morphological markers were used to genotype 270 recombinant inbred lines, and a map including 604 loci (509 SSR, 58 IT-ISJ,
29 SRAP and 8 morphological loci) and 60 linkage groups was constructed. The map spanned 3,140.9 cM with an average interval
of 5.2 cM between two markers, approximately accounting for 70.6% of the cotton genome. Fifty-four of 60 linkage groups were
ordered into 26 chromosomes. Multiple QTL mapping was used to identify QTL for fiber quality traits in five environments,
and thirteen QTL were detected. These QTL included four for fiber length (FL), two for fiber strength (FS), two for fiber fineness (FF), three for fiber length uniformity (FU), and two for fiber elongation (FE), respectively. Each QTL explained between 7.4 and 43.1% of phenotypic variance. Five out of thirteen QTL (FL1 and FU1 on chromosome 6, FL2, FU2 and FF1 on chromosome7) were detected in five environments, and they explained more than 20% of the phenotypic variance. Eleven QTL
were distributed on A genome, while the other two on D genome. 相似文献
14.
Protoplast-to-plant regeneration in cotton (Gossypium hirsutum L. cv. Coker 312) using feeder layers
Summary We report the regeneration of protoplasts isolated from two embryogenic cell lines of Gossypium hirsutum L. cv. Coker 312 initiated from hypocotylderived callus. Protoplasts plated on cellulose nitrate filters and placed over feeder layers formed embryogenic callus from which plants were regenerated. Plating efficiency up to 12.8% depended upon the cell line. Addition of phytohormones to the protoplast medium had no stimulating effect on plating efficiency. The influence of feeder cells and conditioned medium on plating efficiency was significantly different for the two cell lines.Abbreviations ACM
autoclaved conditioned medium
- AFC
autoclaved feeder cells
- BM
basic medium
- BM+
basic medium with phytohormones
- CM
non-autoclaved conditioned medium
- FC
non-autoclaved feeder cells
- FDA
fluorescein diacetate
- MM
maturation medium
- NAA
1-naphtaleneacetic acid
- PCM
protoplast culture medium
- PCM+
protoplast culture medium with phytohormones
- SC
settled cells
- 2,4-D
2,4-dichlorophenoxyacetic acid
- 6-BAP
6-benzylamino purine 相似文献
15.
Dalton R. Gossett Eddie P. Millhollon M. Cran Lucas Stephen W. Banks Marye-Michelle Marney 《Plant cell reports》1994,13(9):498-503
Summary To determine NaCl effects on callus growth and antioxidant activity, callus of a salt-tolerant and a salt-sensitive cultivar of cotton was grown on media amended with 0, 75, and 150 mM NaCl. Callus of the salt-tolerant cultivar, Acala 1517-8 8, grown at 150 mM NaCl, showed significant increases in superoxide dismutase, catalase, ascorbate peroxidase, peroxidase and glutathione reductase activities compared to callus tissue grown at 0 mM NaCl. In contrast, callus tissue of the salt-sensitive cultivar, Deltapine 50, grown at 0, 75, and 150 mM NaCl, showed no difference in the activities of these enzymes. At the 150 mM NaCl treatment, peroxidase was the only antioxidant enzyme from Deltapine 50 with an activity as high as that observed in Acala 1517-88. The NaCl-induced increase in the activity of these enzymes in Acala 1517-88 indicates that callus tissue from the more salt-tolerant cultivar has a higher capacity for scavenging and dismutating superoxide, an increased ability to decompose H2O2, and a more active ascorbate-glutathione cycle when grown on media amended with NaCl. 相似文献
16.
17.
Regeneration of Gossypium hirsutum and G. barbadense from shoot apex tissues for transformation 总被引:1,自引:0,他引:1
Jean Gould Sharon Banister Osamu Hasegawa Michal Fahima Roberta H. Smith 《Plant cell reports》1991,10(1):12-16
A method of regenerating cotton plants from the shoot apical meristem of seedlings was developed for use with particle gun and Agrobacterium-mediated transformation. This method was developed to circumvent the problems of genotype restriction and chromosomal damage frequently encountered in cotton regeneration in tissue culture through somatic embryogenesis. In this procedure, the cells of the shoot meristem are targeted for transformation. Normal and fertile plants of Gossypium barbadense Pima S-6, and 19 cultivars of G. hirsutum were regenerated using this method. Shoot regeneration from these tissues was direct and relatively rapid. A MS based, hormone-free medium could be used with all the varieties tested.This project was funded by grants from Cotton Incorporated, Nisshinbo Industries, and a grant from the Texas Agricultural Experiment Station to RHS. Texas Agricultural Experiment Station Technical Article TA-25667. 相似文献
18.
Zhang J Cai L Cheng J Mao H Fan X Meng Z Chan KM Zhang H Qi J Ji L Hong Y 《Transgenic research》2008,17(2):293-306
While genetically modified upland cotton (Gossypium hirsutum L.) varieties are ranked among the most successful genetically modified organisms (GMO), there is little knowledge on transgene
integration in the cotton genome, partly because of the difficulty in obtaining large numbers of transgenic plants. In this
study, we analyzed 139 independently derived T0 transgenic cotton plants transformed by Agrobacterium tumefaciens strain AGL1 carrying a binary plasmid pPZP-GFP. It was found by PCR that as many as 31% of the plants had integration of
vector backbone sequences. Of the 110 plants with good genomic Southern blot results, 37% had integration of a single T-DNA,
24% had two T-DNA copies and 39% had three or more copies. Multiple copies of the T-DNA existed either as repeats in complex
loci or unlinked loci. Our further analysis of two T1 populations showed that segregants with a single T-DNA and no vector
sequence could be obtained from T0 plants having multiple T-DNA copies and vector sequence. Out of the 57 T-DNA/T-DNA junctions
cloned from complex loci, 27 had canonical T-DNA tandem repeats, the rest (30) had deletions to T-DNAs or had inclusion of
vector sequences. Overlapping micro-homology was present for most of the T-DNA/T-DNA junctions (38/57). Right border (RB)
ends of the T-DNA were precise while most left border (LB) ends (64%) had truncations to internal border sequences. Sequencing
of collinear vector integration outside LB in 33 plants gave evidence that collinear vector sequence was determined in agrobacterium
culture. Among the 130 plants with characterized flanking sequences, 12% had the transgene integrated into coding sequences,
12% into repetitive sequences, 7% into rDNAs. Interestingly, 7% had the transgene integrated into chloroplast derived sequences.
Nucleotide sequence comparison of target sites in cotton genome before and after T-DNA integration revealed overlapping microhomology
between target sites and the T-DNA (8/8), deletions to cotton genome in most cases studied (7/8) and some also had filler
sequences (3/8). This information on T-DNA integration in cotton will facilitate functional genomic studies and further crop
improvement. 相似文献
19.
Transgenic Bt cotton NewCott 33B and transgenic tfd A cotton TFD were chosen to evaluate pollen dispersal frequency and distance of transgenic cotton (Gossypium hirsutum L.) in the Huanghe Valley Cotton-producing Zone, China. The objective was to evaluate the efficacy of biosafety procedures
used to reduce pollen movement. A field test plot of transgenic cotton (6×6 m) was planted in the middle of a nontransgenic
field measuring 210×210 m. The results indicated that the pollen of Bt cotton or tfd A cotton could be dispersed into the environment. Out-crossing was highest within the central test plot where progeny from
nontransgenic plants, immediately adjacent to transgenic plants, had resistant plant progeny at frequencies up to 10.48%.
Dispersal frequency decreased significantly and exponentially as dispersal distance increased. The flow frequency and distance
of tfd A and Bt genes were similar, but the pollen-mediated gene flow of tfd A cotton was higher and further to the transgenic block than that of Bt cotton (χ2 = 11.712, 1 degree of freedom, p<0.001). For the tfd A gene, out-crossing ranged from 10.13% at 1 m to 0.04% at 50 m from the transgenic plants. For the Bt gene, out-crossing ranged from 8.16% at 1 m to 0.08% at 20 m from the transgenic plants. These data were fit to a power curve
model: y=10.1321x
−1.4133 with a correlation coefficient of 0.999, and y=8.0031x
−1.483 with a correlation coefficient of 0.998, respectively. In this experiment, the farthest distance of pollen dispersal from
transgenic cotton was 50 m. These results indicate that a 60-m buffer zone would serve to limit dispersal of transgenic pollen
from small-scale field tests. 相似文献
20.
Organization,inheritance and expression of acetohydroxyacid synthase genes in the cotton allotetraploid Gossypium hirsutum 总被引:1,自引:0,他引:1
John W. Grula Richard L. Hudspeth Susan L. Hobbs David M. Anderson 《Plant molecular biology》1995,28(5):837-846
The acetohydroxyacid synthase (AHAS) gene family of the cotton AD allotetraploid Gossypium hirsutum has been cloned and characterized. We have identified six different AHAS genes from an analysis of genomic clones and Southern blots of genomic DNA. Four of the six genes are organized as tandem pairs, in which the genes are separated by only 2–3 kb. Conservation of restriction fragment length polymorphisms between G. hirsutum and A-genome and D-genome-containing diploid cottons was sufficient to assign the single genes in clones A5 and A19 to the A and D subgenomes, respectively. Each diploid genome has one tandem pair, but in these cases we could not make specific subgenomic assignments. DNA and deduced amino acid sequences were determined for the A5 and A19 genes, and an AHAS cDNA clone isolated from a leaflibrary. The sequence of the A19 gene matches that of the cDNA clone, while the A5 gene is 97.8% similar. The four genes comprising the tandem pairs are much less similar to the cDNA clone. The deduced amino acid sequences of the mature polypeptides encoded by the A5 and A19 genes are collinear with the housekeeping forms of AHAS from Arabidopsis thaliana, Nicotiana tabacum and Brassica napus. The constitutive expression of A5 and A19 was confirmed with RNase protection assays and northern blots. We conclude that these genes encode the main house-keeping froms of AHAS in G. hirsutum. Among the four AHAS genes comprising the two tandem pairs, at least two are functional. These genes exhibit either low-level constitutive expression (one or both of the downstream genes of each pair), or highly specific expression in reproductive tissue (one or both of the upstream genes of each pair). The AHAS gene family of G. hirsutum is more complex than that of other plants so far examined. 相似文献