CHLOROPLASTS OF MONOPLOID AND DIPLOID OENOTHERA HOOKERI

Comparison of leaves from diploid Oenothera hookeri plants with leaves of their monoploid derivatives revealed the following differences: the leaves of the diploids are almost twice as thick as those of the monoploids; the diploids have only one-fifth as many cells per unit leaf area as do the monoploids; leaf tissue of the diploids has an average of 36.3 chloroplasts per cell compared with 27.4 for the monoploids; chloroplasts of the diploids average 4.68 μm in length, those of the monoploids, 3.74 μm; chloroplasts from the diploids contain about seven times more chlorophyll than those of the monoploids and have an average of seven thylakoids per granum compared with an average of only three for the monoploids; the average chlorophyll a/b ratio for the diploid chloroplasts is 2.98 compared with 3.48 for the monoploids.     

Indifference of potato anther culture to colchicine and genetic similarity among anther-derived monoploid regenerants determined by RAPD analysis

Effects of colchicine on androgenesis of diploid potato (Solanum phureja Juz. & Buk.) and ploidy of anther-derived plants were examined in three experiments. In the first, no significant difference was found for mean embryos per anther of an interspecific potato clone after application of five colchicine treatments (0, 25, 50, 100 and 200 mg l^-1) for 24 h to freshly excised anthers containing late uninucleate microspores. The same colchicine treatments were applied to six hybrid potato families in the second experiment. Families differed for number of embryos per anther and embryo regeneration frequency; however, androgenic response did not differ significantly among colchicine treatments. The 312 regenerated plants included 233 (75%) monoploids. The third experiment examined durations (0, 90 s vacuum infiltration, 24, 48 and 72 h) of high colchicine treatment (200 mg l^-1) on anther culture of seedlings representing one family. Mean embryos per anther, though not statistically significant, ranged from 0.96 to 1.90 for 48 h colchicine and 90 s vacuum infiltration, respectively. There were 126 plants regenerated of which 62% were monoploid. Frequency of monoploid plants regenerated from colchicine treatments did not differ significantly. RAPD analysis was conducted on 26 anther-derived monoploids of one family, based on common flasks of origin. The 13 decamer primers revealed 54 polymorphic loci. These were used to characterize the monoploids genetically. From one flask, two pairs of monoploids among six examined were genetically indistinguishable. Examination of a second and third flask revealed, six of seven and three of seven monoploids that were genetically indistinguishable. These data suggest the regeneration of genetic clones within flasks and may indicate the occurrence of secondary embryogenesis during anther culture. This revised version was published online in June 2006 with corrections to the Cover Date.     

Synthesis and Assembly of <Emphasis Type="Italic">Porphyromonas gingivalis</Emphasis> Fimbrial Protein in Potato Tissues

Periodontal disease caused by the gram-negative oral anaerobic bacterium Porphyromonas gingivalis is thought to be initiated by the binding of P. gingivalis fimbrial protein to saliva-coated oral surfaces. To assess whether biologically active fimbrial antigen can be synthesized in edible plants, a cDNA fragment encoding the C-terminal binding portion of P. gingivalis fimbrial protein, fimA (amino acids 266–337), was cloned behind the mannopine synthase promoter in plant expression vector pPCV701. The plasmid was transferred into potato (Solanum tuberosum) leaf cells by Agrobacterium tumefaciens in vivo transformation methods. The fimA cDNA fragment was detected in transformed potato leaf genomic DNA by PCR amplification methods. Further, a novel immunoreactive protein band of ~6.5 kDa was detected in boiled transformed potato tuber extracts by acrylamide gel electrophoresis and immunoblot analysis methods using primary antibodies to fimbrillin, a monomeric P. gingivalis fimbrial subunit. Antibodies generated against native P. gingivalis fimbriae detected a dimeric form of bacterial-synthesized recombinant FimA(266–337) protein. Further, a protein band of ~160 kDa was recognized by anti-FimA antibodies in undenatured transformed tuber extracts, suggesting that oligomeric assembly of plant-synthesized FimA may occur in transformed plant cells. Based on immunoblot analysis, the maximum amount of FimA protein synthesized in transformed potato tuber tissues was approximately 0.03% of total soluble tuber protein. Biosynthesis of immunologically detectable FimA protein and assembly of fimbrial antigen subunits into oligomers in transformed potato tuber tissues demonstrate the feasibility of producing native FimA protein in edible plant cells for construction of plant-based oral subunit vaccines against periodontal disease caused by P. gingivalis.     

HAPLOIDY IN HAPLOPAPPUS GRACILIS (N = 2)

Analysis of meiosis in a haploid (monoploid) sporophyte of Haplopappus gracilis (n = 2) showed a nonrandom distribution of chromosomes at anaphase I. Chromosomes A and B were associated at various prophase I stages in 34 % of the microsporocytes. Presumably, this association persisted long enough to disrupt random distribution in a portion of those meiocytes showing such an association. Pollen stainability of 26.5 % in the haploid was in agreement with the normal expectation. However, this number resulted from a nonrandom chromosome distribution. The usual method of predicting fertility of haploids, 1/2ⁿ, is not accurate for monoploids with low chromosome numbers, and more functional methods are proposed.     

Do ploidy level and nuclear genome size and latitude of origin modify the expression of <Emphasis Type="Italic">Phragmites australis</Emphasis> traits and interactions with herbivores?

We studied the relationship between genome size and ploidy level variation and plant traits for the reed grass Phragmites australis. Using a common garden approach on a global collection of populations in Aarhus, Denmark, we investigated the influence of monoploid genome size and ploidy level on the expression of P. australis growth, nutrition and herbivore-defense traits and whether monoploid genome size and ploidy level play different roles in plant trait expression. We found that both monoploid genome size and latitude of origin contributed to variation in traits that we studied for P. australis, with latitude of origin being generally a better predictor of trait values and that ploidy level and its interaction with monoploid genome size and latitude of origin also contributed to trait variation. We also found that for four traits, tetraploids and octoploids had different relationships with the monoploid genome size. While for tetraploids stem height and leaf water content showed a positive relationship with monoploid genome size, octoploids had a negative relationship with monoploid genome size for stem height and no relationship for leaf water content. As genome size within octoploids increased, the number of aphids colonizing leaves decreased, whereas for tetraploids there was a quadratic, though non-significant, relationship. Generally we found that tetraploids were taller, chemically better defended, had a greater number of stems, higher leaf water content, and supported more aphids than octoploids. Our results suggest trade-offs among plant traits mediated by genome size and ploidy with respect to fitness and defense. We also found that the latitude of plant origin is a significant determinant of trait expression suggesting local adaptation. Global climate change may favor some genome size and ploidy variants that can tolerate stressful environments due to greater phenotypic plasticity and to fitness traits that vary with cytotype which may lead to changes in population genome sizes and/or ploidy structure, particularly at species’ range limits.     

Synthesis and assembly of SIVmac Gag p27 capsid protein cholera toxin B subunit fusion protein in transgenic potato

A deoxyribonucleic acid (DNA) fragment encoding the cholera toxin B subunit (CTB) was linked 5′ to the simian immunodeficiency virus (SIV_mac) Gag p27 capsid gene (CTB-Gag). The fusion gene was transferred into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation methods and transformed plants regenerated. The CTB-Gag gene fusion was detected in transformed potato leaf genomic DNA by polymerase chain reaction-mediated DNA amplification. The results of immunoblot analysis with anti-CTB and anti-Gag antibodies verified the synthesis of biologically active CTB-Gag fusion protein in transformed leaf and tuber tissues. Synthesis and assembly of the CTB-Gag fusion protein into oligomeric structures of pentamer size was confirmed by G_M1-ganglioside-enzyme-linked immunosorbent assay (G_M1-ELISA) of transformed potato tuber tissue extracts. The binding of CTB-Gag fusion protein oligomers to intestinal epithelial cell membrane receptors quantified by G_M1-ELISA showed that CTB-Gag fusion protein made up approx 0.016–0.022% of the total soluble tuber protein. The synthesis of CTB-Gag monomers and their assembly into biologically active CTB-Gag fusion protein oligomers in potato tuber tissues provides the opportunity for employment of the carrier and adjuvant properties of CTB for the development of edible plant-based subunit mucosal vaccines for enhanced mucosal immunity against SIV in macaques.     

Colchicine-induced microspore embryogenesis in coffee

A protocol for the induction of androgenesis and plant regeneration from C. arabica cv. Caturra isolated microspores in vitro using colchicine pretreatment has been developed. Microspores were mechanically isolated and then carefully purified. Before colchicine pretreatment, microspores were cultured in a semi-solid medium for further develop and regeneration. Different times of colchicine exposure as well as different concentrations were tested. The best androgenic response was found when microspores were precultured in 100 mg l^–1 colchicine for 48 h. The microspore developmental stages responsive to colchicine were late-uninucleated and early binucleated pollen. Flow cytometry and morphological analyses revealed that 95% of regenerated plants were dihaploids (2n=2x=22). However, some doubled dihaploid plants (2n=4x=44) were also obtained, suggesting that not only androgenic induction but also chromosome duplication could be expected as result of colchicine exposure of coffee microspores. This report represents a new approach in the coffee pollen culture, as well as a major step forward to the utilization of haploid technology in coffee breeding.     

Monoploids in Zea mays L. following crosses with untreated and X-rayed pollen

Monoploids in Zea mays L. occur spontaneously among individual diploid seedlings. Plants with the gametic chromosome number have also been detected among members of multiple seedlings of maize and numerous other species of angiosperms. Previous reports disclosed that Xirradiation of the pollen successfully stimulated reduced parthenogenesis in some other angiosperms, but the results of X-ray treatment were inconclusive in maize. Therefore, a tester stock of maize homozygous for lg _land gl _l was crossed with pollen from inbred CI3A, carrying the dominant alleles. The pollen was exposed to 0, 1000, 2000, and 4000r units of X rays. chromosome counts were made from root tips of plants exhibiting both recessive phenotypes to establish the frequencies of monoploids in the control and X₁ populations.Monoploids were more abundant among the individual seedlings from crosses with untreated pollen than in the X₁ populations. X irradiation of the pollen is not a feasible method for the induction of monoploids in maize. The X-ray treatments greatly increased the frequency of multiple seedlings, and deficiencies were numerous among them. The members of a set of multiple seedlings were always genetically identical, and no monoploid members occurred. It is concluded that the induced deficiencies caused atypical development resulting in zygotic or embryonic cleavage.Scientific Article No. A2070, Contribution No. 5023 of the Maryland Agricultural Experiment Station, Department of Botany.     

Effects of colchicine on polyploidy induction of Buddleja lindleyana seeds

Buddleja lindleyana Fort. is a garden ornamental plant with great potential for development and also a commonly used medicinal plant. To enrich its germplasm resources, the seeds of B. lindleyana were treated with colchicine solution with concentration gradients of 0.5%, 1.0%, 1.5%, 2.0% and 3.0% for 12-, 24- and 48-h respectively, and the water treatment was set as the control group. The purpose was to explore the effects of colchicine on the germination and mutagenic effect of B. lindleyana seeds at different concentrations and different times, to screen the appropriate combination of mutagenic concentration and time, to provide guidance for the construction of B. lindleyana mutation population in future research. The results were as follows: (1) Colchicine had an inhibitory effect on seed germination and seedling height of B. lindleyana seeds, and the higher the concentration, the more obvious the inhibitory effect. (2) After colchicine treatment, 30 mutant plants showed morphological variations such as leaf malformation, leaf color macular, early leaf bud germination, uneven leaf surface and leaf hyperplasia, among which 3.0%?+?48-h treatment group had great potential to produce yellow-leaf plants. (3) Detection and analysis by flow cytometry revealed that among the 30 morphologically variant plants, there were 22 diploid plants, 3 tetraploid plants, and 5 chimera plants. Among them, tetraploids were mainly from colchicine concentration of 3.0% (2 plants) and 1.5% (1 plant), chimeras were mainly from colchicine concentration of 1.0% (2 plants), 1.5% (1 plant) and 3.0% (2 plants), and the seed soaking time was 48-h. (4) The length and width of guard cells and stomata were significantly different between diploid and tetraploid, and there were significant differences in leaf width and leaf shape index between tetraploid and diploid, but there were no significant differences in leaf length among diploid, tetraploid and chimera. In short, we got tetraploids and chimeras materials which were potentially useful cultivars of B. lindleyana and provided an effective identification method for polyploids of B. lindleyana.

     

Leaf proteomic analysis in cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis>, Crantz) during plant development,from planting of stem cutting to storage root formation

Tuberization in cassava (Manihot esculenta Crantz) occurs simultaneously with plant development, suggesting competition of photoassimilate partitioning between the shoot and the root organs. In potato, which is the most widely studied tuber crop, there is ample evidence suggesting that metabolism and regulatory processes in leaf may have an impact on tuber formation. To search for leaf proteins putatively involved in regulating tuber generation and/or development in cassava, comparative proteomic approaches have been applied to monitor differentially expressed leaf proteins during root transition from fibrous to tuberous. Stringent cross comparison and statistical analysis between two groups with different plant ages using Student’s t test with 95% significance level revealed a number of protein spots whose abundance were significantly altered (P < 0.05) during week 4 to week 8 of growth. Of these, 39 spots were successfully identified by ion trap LC–MS/MS. The proteins span various functional categories from antioxidant and defense, carbohydrate metabolism, cyanogenesis, energy metabolism, miscellaneous and unknown proteins. Results suggested possible metabolic switches in the leaf that may trigger/regulate storage root initiation and growth. This study provides a basis for further functional characterization of differentially expressed leaf proteins, which can help understand how biochemical processes in cassava leaves may be involved in storage root development.     

Somatic chromosome number doubling of selected potato genotypes using callus culture or the colchicine treatment of shoot nodes in vitro

Experiments were carried out to double the somatic cell chromosome numbers of a monoploid and dihaploid of Solanum tuberosum and a genotype of S. circaeifolium subsp. quimense. Colchicine was used in vitro on shoot nodes from which the axillary meristems had been removed. Plants with doubled chromosome numbers were obtained from shoots grown from the tertiary, sub-axillary meristems of all three genotypes. The callus culture of stem and leaf explants was found to produce more shoots with doubled chromosome numbers than the colchicine treatment in the case of the dihaploid and quimense genotypes but no shoots were obtained from callus culture of the monoploid. Fifty-two % of the shoots from the dihaploid and 63% from the quimense clone were ploidy doubled in the case of the best callus culture system. Using a sub-lethal dose of colchicine, the dihaploid yielded 37% ploidy-doubled shoots whereas all the shoots produced from the monoploid were doubled and the quimense clone produced 27% doubled plants. Callus culture was highly dependent upon the type of growth medium and other, unknown, factors. The colchicine treatment, although yielding fewer products, was more reliable for achieving ploidy doubling in selected clones if the number of plants produced is not important.     

Response of potato grown under non-inductive condition paclobutrazol: shoot growth, chlorophyll content, net photosynthesis, assimilate partitioning, tuber yield, quality, and dormancy

The effect of foliar and soil applied paclobutrazol on potato were examined under non-inductive condition in a greenhouse. Single stemmed plants of the cultivar BP1 were grown at 35(±2)/20(±2) °C day/night temperatures, relative humidity of 58%, and a 16 h photoperiod. Twenty-eight days after transplanting paclobutrazol was applied as a foliar spray or soil drench at rates of 0, 45.0, 67.5, and 90.0 mg active ingredient paclobutrazol per plant. Regardless of the method of application paclobutrazol increased chlorophyll a and b contents of the leaf tissue, delayed physiological maturity, and increased tuber fresh mass, dry matter content, specific gravity, dormancy period of the tubers. Paclobutrazol reduced the number of tubers per plant. A significant interaction between rates and methods of paclobutrazol application were observed with respect to plant height and tuber crude protein content. Foliar application gave a higher rate of net photosynthesis than the soil drench. Paclobutrazol significantly reduced total leaf area and increased assimilate partitioning to the tubers. The study clearly showed that paclobutrazol is effective to suppress excessive vegetative growth, favor assimilation to the tubers, increase tuber yield, improve tuber quality and extend tuber dormancy of potato grown in high temperatures and long photoperiods.     

不同种源樟树叶片形态特征及生长差异分析

为了解不同种源樟树叶片形态特征和生长差异,该文以30个种源樟树为研究对象,对其叶长、叶宽、叶柄长、周长、叶面积、长宽比、形态因子、株高、地径等指标进行测定和差异性分析.结果表明:(1)30个种源间叶片性状的变异系数为3.88％～16.14％,显示不同种源樟树叶片形态特征存在显著差异;叶长、叶宽、叶柄长、周长、面积、叶厚...     

Androgenesis in the vine cacti <Emphasis Type="Italic">Selenicereus</Emphasis> and <Emphasis Type="Italic">Hylocereus</Emphasis> (Cactaceae)

Anther culture techniques were applied to develop a methodology for producing haploid vine cactus plants. Anthers with most microspores in the middle uninucleate developmental stage from the tetraploid species Selenicereus megalanthus and the diploid species Hylocereus polyrhizus and H. undatus were cultured on basal MS medium supplemented with picloram and 6-benzyladenine (BA). H. polyrhizus and H. undatus anthers were also cultured on MS medium containing either thidiazuron (TDZ) or 2,4-dichlorophenoxyacetic acid (2,4-D). Pro-embryo development started after three days of culture. A direct androgenic embryo response was achieved in S. megalanthus with and without picloram/BA, while H. polyrhizus exhibited non-regenerative callus formation. Only a single direct androgenic embryo was obtained in H. polyrhizus (with 0.1 mg/l TDZ). H. undatus required a cold pre-treatment of 4°C for 24 h in 0.3 M D-mannitol to produce a response, but only calluses were obtained and they did not regenerate. S. megalanthus and H. polyrhizus embryos converted into plantlets after transfer to MS medium in the light. Rooted plants were acclimatized successfully, and most plants showed normal phenotypes. Flow cytometry and cytological studies revealed monoploid, haploid, dihaploid, and mixoploid plants. This study showed that androgenesis is strongly species- and culture-medium-dependent, thus revealing new perspectives in the genetics and breeding of vine cactus species. To the best of our knowledge, this is the first report on the production of monoploid, haploid and dihaploid plants in Cactaceae.     

Morphological, molecular and cytological analyses of Carica papaya×C. cauliflora interspecific hybrids

 Morphological, molecular and cytological analyses were performed to assess the hybridity of 120 putative interspecific hybrids of Carica papaya L.×C. cauliflora Jacq. In the putative interspecific hybrids the number of main leaf veins was intermediate between the two parents while the hermaphrodite flower sex form and the low vigour were distinctive features of these hybrids. Petiole length, stem diameter, leaf length, leaf width and flower colour were similar to C. papaya, whereas leaf shape, type, serration, venation, petiole hairiness and flower shape were similar to C. cauliflora. Markers generated by the polymerase chain reaction using 72 10-mer primers (random amplified polymorphic DNA) revealed a high level of polymorphism (64%) between C. papaya and C. cauliflora. Seventeen of these primers yielded reliable and easily scorable polymorphic banding patterns that were further screened to reveal hybrids. A range of 1–5 RAPD primers consistently confirmed that all 120 plants were genetic hybrids, with all of them containing at least one band from the male parent. Cytological analysis revealed that 7–48% of the cells in many of the interspecific hybrids were aneuploid suggesting that chromosome elimination was occurring. The frequency of aneuploid cells was negatively associated (r=0.88) with the number of bands from the male parent integrated into the hybrid. Pollen fertility of the hybrids was from 0.5 to 14.0% while C. papaya and C. cauliflora had 88.0–99.0% and 90.0–97.0% fertile pollen, respectively. Received: 28 February 1996 / Accepted: 27 September 1996     

Molecular Phylogeny of Eupatorieae (Asteraceae) Estimated from cpDNA RFLP and its Implication for the Polyploid Origin Hypothesis of the Tribe

Neomirandea (x=17 and 25), Ageratina (x=17) and Sclerolepis (x=15) with the higher chromosome base numbers, and the other includes Mikania (x=17) and the remaining genera with lower chromosome base numbers (x=10–11). However, the monophyly of the former clade is supported with a low bootstrap value. In the latter clade, Mikania (x=17) diverged first, then Stevia (x=11), and finally eight genera with x=10 diverged in succession. This result supports the hypothesis that the genera in the tribe Eupatorieae with x =10 evolved from an ancestor with a higher base number, and the tribe is of polyploid origin. Received 13 September 1999/ Accepted in revised form 20 January 2000     

广西多穗柯叶片性状变异及幼苗生长量研究

多穗柯是一种珍贵天然野生药用植物,可以开发出保健食品色素和天然医用药品,广西的资源较丰富,该研究采集巴马、那坡、德保及田林等4个产地的多穗柯种子进行播种育苗,并跟踪调查测定一年生幼苗的叶片性状及幼苗生长量,并进行相关性分析。结果表明:(1)不同产地间叶片性状及幼苗生长指标均存在不同程度的差异,其中巴马与那坡、德保、田林在叶长、叶宽、叶面积、叶脉间距、叶鲜重、叶片干物质含量、叶片组织密度等叶片性状上的差异均达到显著水平,在株高、地径、单株干重、主根长、单株根干重及单株叶干重等生长指标上亦存在显著差异,且生长量是后3个产地的1~2倍;通过比较各产地的叶片保水力及植株净生长量,巴马的多穗柯植株耐旱性及生长速度优于其他三地。综合各性状表现,认为巴马的多穗柯苗期表现比较好,生长速度快,长势好,抗旱性较强,可作为多穗柯优良种源的初步选择。(2)8月份是多穗柯株高、地径的生长高峰期,建议此时应加强肥水管理,调节适宜的水肥光热条件,尽量延长幼苗的快速生长时间,以获得苗木的最大累积生长量。(3)叶片性状与幼苗生长量的相关性分析结果显示,叶面积与株高、地径、单株干重、单株根干重以及单株叶干重等呈极显著正相关,叶脉间距、叶绿素相对含量(SPAD)与株高、单株干重呈显著或极显著正相关,比叶面积与株高、地径呈显著负相关。因此,在以后的优株表型选择中,要优先考虑叶子大、叶脉间距宽、中老熟叶片叶色浓绿的植株。该研究结果为多穗柯优良种质资源的早期筛选提供了一定的依据。     

Effects of Benzyladenine and Naphthalene Acetic Acid on Growth and Camptothecin Accumulation in Camptotheca acuminata Seedlings

The effects of the cytokinin benzyladenine (BA) and the auxin naphthalene acetic acid (NAA) on Camptotheca acuminata Decaisne growth and camptothecin (CPT) accumulation (leaf CPT concentration and total leaf CPT yield) were studied in a hydroponic culture system for three weeks. Increasing BA concentrations from 0 to 3 mg l^–1 in growth medium decreased plant height, stem weight, and leaf weight but increased root weight. High BA levels (1 and 3 mg l^–1) increased leaf CPT concentration (% of dry weight), whereas BA applications had no effect on total leaf CPT yield, the product of leaf CPT concentration and total leaf dry weight per seedling. There was a positive correlation between root weight and leaf CPT concentration under BA treatments. NAA supplementations (from 0.5 to 4 mg l^–1) to growth medium reduced plant height, leaf number, leaf length, specific leaf weight, plant weight, stem weight, and leaf weight compared with the NAA control. Meanwhile, there were no differences in plant height, leaf length, and specific leaf weight among the NAA supplementations. NAA applications had no effect on leaf CPT concentration and NAA applications decreased total leaf CPT yield. There were negative correlations between leaf number and leaf CPT concentration, leaf length and leaf CPT concentration under NAA treatments. Our results suggest that BA applications from 0.3 to 3 mg l^–1 are not helpful for achieving high total leaf CPT yield and NAA applications from 0.5 to 4 mg l^–1 decrease total leaf CPT yield.     

Heterologous Expression of a Gibberellin 2-Oxidase Gene from <Emphasis Type="BoldItalic">Arabidopsis thaliana</Emphasis> Enhanced the Photosynthesis Capacity in <Emphasis Type="BoldItalic">Brassica napus</Emphasis> L.

Gibberellins (GAs) are endogenous hormones that play an important role in regulating plant stature by increasing cell division and promoting seed germination. The GA2-oxidase gene from Arabidopsis thaliana (AtGA2ox8) was introduced into Brassica napus L. by Agrobacterium-mediated floral-dip transformation with the aim of decreasing the amount of bioactive GA and hence reduced the plant height. As anticipated, the transgenic plant exhibited dwarf phenotype. Importantly, compared with the wild type, the transgenic plants had delayed the seed germination, increased the chlorophyll content (28.7–36.3%) and photosynthesis capacity (14.3–18.7%) in a single leaf. At the same time, the photosynthesis capacity of the whole plants was significantly enhanced (35.7–48.6%) due to the extra leaves and branches.     

Callus growth,tumour development and polyploidization in the tetraploid potato cultivar bintje

Various combinations of exogenously applied growth regulators supported callus growth to different extents in cultures of leaf and tuber explants of the tetraploid potato cv. Bintje (Solanum tuberosum L.). Various strains of Agrobacterium tumefaciens differing in induction of endogenous hormonal activities, induced tumours of varying habitus and frequency on tuber dics. The ploidy levels of cells from tissues were determined by means of flow cytometry. In the original explants, varying degrees of polyploidization were observed. Moreover, upon ageing of explanted leaf tissues, rapid endopolyploidization occurred. The DNA histograms of cell populations of calli or tumours were similar, irrespective of hormonal regime and explant source. Within 1 month of growth, a stable pattern of polyploidization was reached, the DNA histograms resembling the patterns found in tuber tissues and in aged leaf explants, cultured in the absence of plant growth regulatory substances. The factors influencing genetic stability, are attributed to predominant genotype effects.