Improvement of green plant regeneration by manipulation of anther culture induction medium of hexaploid wheat

Anthers of three hexaploid wheat (Triticum aestivum L.) genotypes with high frequencies of albino regenerants in anther culture were compared to DH after inoculation on medium supplemented with ficoll, colchicine or maltose separately, pair-wise or combined, in an attempt to increase green plant regeneration. Maltose treatment produced more green regenerated plants than sucrose for all of the genotypes. The three chemicals combined in anther medium either reduced green plant regeneration or did not yield significantly different numbers of green regenerated plants compared to the maltose treatment. With DH fewer embryo-like structures (ELS) were obtained per 100 cultured anthers on all medium containing colchicine but greater frequencies of green plants per 100 ELS were obtained. It appeared that the increase in green regenerated plants per 100 ELS was due to a better quality of embryos that were capable of regenerating into green rather than albino plantlets. Smaller increases in green plants per 100 ELS were observed in ICR 4 and V-15 on colchicine containing medium compared to DH. Genotypic differences in anther culture response were observed for ELS per 100 cultured anthers (increased for V-37, decreased for DH and approx. the same for ICR 4 and V-15 in medium with all three chemicals compared to the sucrose control).     

Physiological studies on Phymatotrichum omnivorum. IX. Synthesis of indole acetic acid in vitro.

The cotton root rot fungus, Phymatotrichum omnivorum, synthesized indole acetic acid (IAA) in culture medium containing tryptophan as the only source of nitrogen. Synthesis of IAA was not affected by illumination, and maximum amounts of the auxin were produced at 30 days. The optimum hydrogen ion concentration and temperature for IAA production were 6.5 and 28 degrees C, respectively. Indole ethanol (IEA) was also detected in tryptophan media, primarily during the active growth of the organism.     

Gibberellic acid and indole acetic acid compete in ethylene promoted abscission

Summary Gibberellic acid inhibition of betacyanin biosynthesis has been studied in Amaranthus caudatus L. using the pigment precursors L-tyrosine and L-dihydroxyphenylalanine. Precursors fed to gibberellic acid (GA) treated seedlings completely recovered betacyanin synthesis while the GA induced growth enhancement remained unaltered. Inhibition of betacyanin biosynthesis by GA is related to depletion of metabolites/amino acids and their diversion to support changed pattern of metabolism leading to growth.Abbreviations GA gibberellic acid - L-DOPA L-dihydroxyphenylalanine - Pr phytochrome red absorbing form - Pfr phytochrome, far-red absorbing form     

Effects of benzisoxazole and benzisothiazole on tomato plant regeneration in vitro

The effects of two synthetic auxins, BOA and BIA, on plant regeneration in vitro have been studied on explants of tomato cotyledons. The activity of these substances on cell elongation has also been tested on pea stem segments. It has been found that BOA is particularly effective in inducing the formation of shoots but has a weak activity on cell elongation, while BIA, which is more effective in inducing cell elongation, is less active in morphogenesis. It is concluded that (1) the two activities are not related to each other, (2) the receptors involved in the two processes are probably different, (3) thechemical structure of the auxin may be an important factor in organogenetic processes.Abbreviations BIA 1,2-benzisothiazole-3-acetic acid - BOA 1,2-benzisoxazole-3-acetic acid - IAA indoleacetic acid - MS Murashige & Skoog medium     

Effects of different 2,4-D concentrations on the cytogenetic behaviour of plant cells culturedin vitro

Cytogenetic effects of different concentrations of 2,4-D on suspension cultures ofVicia hajastana Grossh. andHaplopappus gracilis (Nutt.) Gray were studied using a simple, chemically defined medium. InVicia, the frequency of dicentric chromosomes showed a negative association with the 2,4-D concentration. In both the species, the frequencies of anaphase bridges were negatively associated, while those of the diploid cells showed a positive association with the 2,4-D concentration in the culture medium. A decrease in the frequency of diploid cells was observed in the cultures transferred to a lower concentration of 2,4-D. This decrease was transient inVicia, while inH. gracilis a new equilibrium at a lower level was established. Both endoreduplication and selection appear to be involved in the achievement of the lower equilibrium inH. gracilis, although selection appears to be the major factor.     

Optimization of indole acetic acid production by isolated bacteria from Stevia rebaudiana rhizosphere and its effects on plant growth

The ability to synthesize Indole-3-acetic acid (IAA) is widely associated with the plant growth promoting rhizobacteria (PGPR). The present work deals with isolation and characterization of such bacteria from the rhizosphere of medicinal plant Stevia rebaudiana and optimization of IAA production from its isolates. The optimization of IAA production was carried out at different pH and temperature with varied carbon and nitrogen sources of culture media. Out of different isolates obtained, three of them were screened as efficient PGPRs on the basis of different plant growth promoting attributes. Isolates CA1001 and CA2004 showed better production of IAA at pH 9 (91.7?µg?ml^?1) and at temperature 37?°C (81.7?µg?ml^?1). Dextrose (1%) was found to be the best carbon source for isolate CA1001 with 104?µg?ml^?1 IAA production. Isolate CA 2004 showed best production of IAA 36?µg?ml^?1 and 34?µg?ml^?1 at 1.5% and 1% Beef extract as nitrogen source respectively. Isolate CA 1001 showed 32?µg?ml^?1 IAA production at 0.5% nicotinic acid concentration. From the current study, CA1001 and CA2004 emerged as noble alternatives for IAA production further which also resulted in root and shoot biomass generation in crop plants, hence can be further used as bio-inoculants for plant growth promotion.     

Effects of incubation temperature on microspore callus production and plant regeneration in wheat anther cultures

Anthers of wheat cultivars Orofen and Pitic 62 were incubated for 8 days at 15, 20, 25, 30, 35 and 40°C before transfer to 25°C. Compared with anthers cultured at 25°C constantly, anthers treated at 30°C produced 40% more microspore callus and green plants in both cultivars whereas those treated at 35°C produced 2–3 fold more green plants. Treatment at 40°C was deleterious. Possible modes of action of high temperature on callus production and albinism were discussed.     

Apical dominance and the levels of indole acetic acid in Phaseolus lateral buds

Combined gas chromatography-mass spectrometry procedures have been used to establish that the indole acetic acid levels of lateral buds from Phaseolus seedlings rise following removal of the shoot apex.Abbreviations GC gas chromatograph - GC-MS combined gas liquid chromatography mass spectrometry - bis-TMS bis-trimethylsilyl - IAA indole acetic acid - MPM multiple-peak monitoring - MS mass spectrometer - GLC gas liquid chromatography - TLC thin-layer chromatography     

Stomatal responses of Vicia faba L. to indole acetic acid and abscisic acid

Evidence is presented that stomata in isolated epidermal peelsof Vicia faba L. open in darkness in response to the externalpresence of indole acetic acid (IAA) in the incubation medium.The effect of IAA is found to be overcome completely in thepresence of either TRIS or MES buffers. In the absence of buffer,V. faba stomata are shown to be influenced by IAA in a concentration-dependenttrend which reached a maximum at an [IAA] of 10^–3 molm^–3. Further investigations reveal that stomata in thisspecies can be shown to respond to the presence of IAA and anotherphytohormone, abscisic acid (ABA). IAA and ABA are demonstratedto be antagonistic in their effects provided the incubationconditions are suitable. The data are discussed in relationto stomatal responses of other species in different treatmentconditions. Recommendations are made with respect to standardizationof incubation media during epidermal peel experiments. Key words: Vicia faba, stomata, indole acetic acid, abscisic acid, buffers     

不同浓度臭氧水对牙周致病菌的体外抑菌作用

目的 探讨不同浓度臭氧水对4种牙周致病菌的体外抑菌效果。方法 采用定量悬液法,分别用1.02 ppm、2.03 ppm和3.88 ppm三种浓度臭氧水,0 ppm无臭氧水溶液（阴性对照组）及3%双氧水（阳性对照组）,对常见的4种牙周致病菌牙龈卟啉单胞菌（Porphyromonas gingivalis）、具核梭杆菌（Fusobacterium nucleatum）、中间普氏菌（Prevotella intermedia）和黏性放线菌（Actinomyces viscosus）分别作用30 s、60 s后,观察不同作用时间下3种不同浓度臭氧水的抑菌效果。结果 对P. gingivalis、F. nucleatum、P. intermedia和A. viscosus作用30 s和60 s,与阴性对照组比较,1.02 ppm、2.03 ppm和3.88 ppm三种浓度臭氧水均具有明显的抑菌效果,3.88 ppm臭氧水与3%双氧水对4种细菌作用60 s的抑菌效果相同。 结论 实验所用的3种不同浓度臭氧水对牙周致病菌P. gingivalis、F. nucleatum、P. intermedia和A. viscosus均有抑菌作用,浓度越高,抑菌效果越好,臭氧水应用于牙龈牙周疾病临床防治有一定的价值和可行性。     

Preussia sp. BSL-10 producing nitric oxide,gibberellins, and indole acetic acid and improving rice plant growth

Preussia sp. have been least known to improve plant growth and produce phytohormones. The current study investigated the production of nitric oxide (NO), indole-3-acetic acid (IAA), and gibberellins (GA₄, GA₇, GA₁₅, and GA₅₃) by a novel endophytic-fungal strain Preussia sp. BSL-10 using advanced chromatographic and spectroscopic techniques. Production of these phytohormones were validated by RT–PCR analysis, which indicated the expression of genes encoding tryptophan synthase (TRP), indole-3-acetamide hydrolase (IAAH), tryptophan-2-monooxygenase (IAAM), aldehyde dehydrogenase (ALD), GA₄ desaturase (DES), geranylgeranyl-diphosphate synthase (GGS2), ent-desaturase oxidase (P450-4), GA₁₄ synthase (P450-1) and nitrite reductase (NIRK/NIRS), cytochrome P450 (P450nor), nitrate reductase (NR), NOS-like (NOL), and nitric oxide reductase (QNOR/CNOR). In plant growth-promoting effects, the inoculation of Preussia sp. BSL-10 significantly increased the growth of dwarf mutant Waito-C and wild-type rice cultivars. In conclusion, utilizing new endophytic with the ability to produce NO, IAA, and gibberellins can be used to promote growth and yield of marginalized crops.     

Effects of acetic acid treatment on plant chromosome structures analyzed by atomic force microscopy

Acetic acid treatment has been frequently used to remove cellular contaminants from plant chromosome samples for structural analyses by scanning electron microscopy and atomic force microscopy (AFM). We evaluated the effects of various concentrations of acetic acid treatments on barley chromosome structures by using AFM. The long-term 45% acetic acid treatment significantly damaged the chromosome structures, although the treatment effectively removed the cellular contaminants. On the other hand, the treatment with 15% acetic acid could not obtain sufficiently clean chromosome samples and the chromosome surface structures could not be observed. In contrast, we obtained clean chromosome preparation without severe damage by using an intermediate concentration (30%) of acetic acid treatment. In the centromeric region, we could observe fiber structures with a width of 100 nm, which were composed of ca. 50-nm granules and aligned to the axes of chromosomes. Thus, AFM analysis of chromosomes appropriately treated with acetic acid will provide important insights into the organization of higher-order structures of plant chromosomes.     

Thidiazuron promotes in vitro regeneration of wheat and barley

Summary Thidiazuron (TDZ) is a substituted phenylurea which has been shown to be an efficacious regulator of in vitro morphogenesis of many dicot plant species. However, information regarding the effect of TDZ on in vitro regeneration of monocot species is limited. We investigated the effects of TDZ on in vitro regeneration of barley (Hordeum vulgare L.) and wheat (Triticum aestivum L.) and found that it promoted shoot regeneration from callus in these two important cereal species. Plant regeneration from calluses derived from immature embryo culture of barley and wheat was observed in regeneration media with a wide range of TDZ concentrations (0.045–45 μM). Shoot regeneration from barley calluses was the highest (38.3% for cv. Golden Promise) at 4.5 μM (1 mg l⁻¹) TDZ, while the optimal TDZ level for wheat regeneration seemed to be 0.9 μM (0.2 mg l⁻¹) (87% for cv. Bob White and 49.4% for cv. Hi Line). Roots developed normally when the regenerated wheat and barley shoots from TDZ-containing media were transferred to the rooting medium. Comparison with other plant growth regulators commonly used in wheat and barley regeneration media suggested that TDZ was among the best for in vitro regeneration of wheat and barley. Both authors contributed equally     

水稻种子内生细菌多样性及其分泌植物生长素能力的测定

[目的]探讨水稻种子内生细菌的多样性并测定其分泌IAA能力.[方法]采用传统的可培养方法分离水稻种子内生细菌,并通过16S Rrna基因序列分析初步确定分离菌株的系统发育地位,利用比色法进一步对不同种类菌株产植物生长素(IAA)能力进行定性、定量检测.[结果]共分离纯化获得66株内生细菌菌株,分属于5个类群的15个属26个种.以26株细菌为代表对其进行分泌生长素(IAA)能力的定性及定量测定,共发现19株细菌可分泌生长素或其类似物,其中Z10、Z17、Z14和Z20 4株内生细菌具较强的分泌植物生长素能力.[讨论]分离得到的内生细菌表现了水稻种子内生细菌的多样性,其中某些细菌对植物有一定的促生功能.     

Genotype and age effect on in vitro embryo rescue of bread wheat x hexaploid triticale hybrids

The in vitro embryo rescue response of hybrid embryos of different ages and sizes of 16 bread wheat x hexaploid triticale crosses was studied. Response was dependent on the parental wheat genotype. However, the triticale genotype did not seem to affect the response of in vitro embryo rescue. Optimum age of the embryo for better recovery of complete plantlets coincided with onset of the drying of hybrid caryopses in the field. Embryo size was positively correlated with embryo germination and recovery of plantlets and negatively correlated with callus development. The best embryo germination (55.55%) and plantlet recovery (50%) were recorded in the hybrid HD 2380 wheat x JNIT 173 triticale.     

Two pathways of plant regeneration in wheat anther culture

The anthers of 10 Polish winter wheat (Triticum aestivum L.) cultivars were used for the induction of androgenesis and plant regeneration. The highest rate of callus induction (9.1%) and green plant production (0.8%) was obtained with the cultivar Apollo that was chosen for histological analysis. The first androgenic division was symmetrical and occurred after 3 weeks of culture. Further divisions of newly formed cells gave rise to multicellular structures which followed two developmental pathways: callus production or direct embryo formation. Plant regeneration was observed in both pathways. Chromosome counting of plantlets regenerated showed that haploid metaphases 2n=3x=21 were the most frequent.     

Involvement of Azopirillum brasilense plasmid DNA in the productio of indole acetic acid

Indole acetic acid (IAA) production in Azospirillum brasilense strain Sp245 is controlled by a 85 MDa plasmid naturally present in this bacterium. In the presence of L-tryptophan, anthranilic acid production and almost no IAA production occurs in a derivative strain harbouring a Tn5-Mob insertion in the 85 MDa plasmid. Agrobacterium tumefaciens strain GM19023, upon transfer of Tn5-Mob labelled 85 MDa plasmid of A. brasilense Sp245, gains the ability to produce anthranilic acid.