Field experiments with Glycine max (L.) Merr. cv. Ludou 11 and Ludou 4 were conducted to evaluate changes in photosynthetic rate, antioxidative enzyme activity, soluble protein, chlorophyll (Chl) and carotenoid (Car) contents in relation to leaf senescence during seed filling period. Photosynthetic rate, soluble protein content, catalase and peroxidase activities were the highest at 25 days after flowering (DAF). Chl a, Chl b and Car contents reached the maximum at 15 DAF and rapidly decreased after 33 DAF.

Two small cationic peptide fractions (5 kDa) were isolated from dry and germinated seeds of wheat, named WAP and GWAP, respectively. The antifungal and antibacterial activities of the peptides were analyzed using disk diffusion and turbidity measurement assays. The peptides in vitro exhibited effective antifungal activity against four plant pathogenic fungi at minimum concentration of 15 μg(protein) cm^-3. Their antimicrobial activity was negatively affected by the presence of 5 mM CaCl₂. The peptides were less effective against Gram-negative bacterium Erwinia carotovora subsp. carotovora, but they demonstrated inhibitory activity against Gram-positive bacterium Clavibacter michiganensis subsp. sepedonicus. The antimicrobial activity of GWAP was more effective than WAP.

The germination of lentil seeds was gradually reduced when seeds were exposed to temperature of 30 or 40 °C, either alone or combined with 0.1, 0.2 or 0.3 M NaCl or 34.1 % (m/v) PEG 8000, during 6 -12 h imbibition. [³⁵S]-methionine incorporation in 12 h imbibed lentil axes also decreased with increasing NaCl concentration at 20 and 40 °C, whereas at 30 °C only 0.3 M NaCl treatment partially inhibited protein synthesis. An analysis of newly synthesized proteins by 1-D SDS PAGE, showed that the expression of most polypeptides decreased following increasing stress. Among these, low molecular mass heat-shock proteins declining, higher in 40 °C treated axes than those treated at 30 °C, supports the hypothesis that at this temperature maximal level of expression of these proteins was achieved.

A protocol for somatic embryogenesis in Azadirachta indica A Juss. has been standardized using in vivo leaflets. Experiments were carried out to examine the effect of various auxins, cytokinins, sucrose, inorganic and organic salts on subsequent somatic embryo induction and maturation. Embryogenic calli were induced on Murashige and Skoog (MS) medium supplemented with 1.5 mg dm^-3 kinetin and 1.5 mg dm^-3 indole-3-acetic acid and subsequently all the stages of somatic embryo development (globular, cordate, torpedo and cotyledonary) were observed. Maturation of these embryos was accomplished with the same growth regulators after three subcultures. The histological study of somatic embryos showed resembles to zygotic embryos. The matured somatic embryos were transferred onto half strength MS-medium devoid of growth regulators for their germination (82 %). Plantlets were acclimatized in the field with a survival rate of 80-83.5 %.

The relationships among 20 samples belonging to 6 subspecies of Vicia sativa based on the variability of seed storage proteins and esterase isozyme electrophoretic patterns was discussed in relation to variation in their morphology and chromosome characters. Electrophoretic protein profiles of different accessions of the same subspecies showed identical (e.g. macrocarpa and cordata) or similar (e.g. amphicarpa) patterns, confirming the stablity of seed storage proteins within these subspecies. However, considerable variation of protein patterns were observed within accessions of both nigra and sativa subspecies, which could be correlated to different geographical origins. Esterase pattern revealed a sharp distinction for each subspecies according to the number and loci of allelic bands. The dendrogram delimited the subspecies incisa and sativa as two separate groups, while the other subspecies were grouped together in another group.

A Pseudomonas strain MRS16 inhibited growth of different pathogenic fungi (Aspergillus sp., Fusarium oxysporum, Pythium aphanidermatum and Rhizoctonia solani) in vitro. Larger inhibition zones were obtained on nutrient agar and King's B media compared to potato dextrose agar and pigment production media. Mutants altered in production of fluorescent pigment were derived by nitrosoguanidine mutagenesis. The pigment overproducer mutant MRS16M-1 was more inhibitory whereas nonproducer mutant MRS16M-5 was less inhibitory than parent strain on nutrient agar medium. Addition of iron (100 µM FeCl₃) in the medium decreased inhibition of fungal growth, suggesting the involvement of siderophores and other antifungal secondary metabolites. Seed bacterization of two cultivars of chickpea (Cicer arietinum cvs. H8618 and C235) differing in susceptibility to wilt caused initial root and shoot stunting at 5 d of growth followed by proliferation of secondary root growth at 10 d. Coinoculation of chickpea with Pseudomonas strain MRS16 or mutants and Rhizobium sp. Cicer strain Ca181 enhanced nodulation, nitrogen fixation and plant dry mass as compared to single inoculation with Rhizobium strain under sterile conditions.

Anthocyanin accumulation and expression pattern of anthocyanin biosynthesis genes were investigated in developing coleoptiles of wheat (Triticum aestivum L.). In epidermal cell layers of the growing coleoptiles of cv. Hope, anthocyanins started to accumulate between day 2 and 3 after germination, reached their maximum on day 6 and then decreased while another cultivar, Chinese Spring (CS) did not accumulate anthocyanin pigments. None of the six anthocyanin biosynthesis genes was upregulated in coleoptiles of both cvs. grown in the dark, whereas all genes were activated by light in coleoptiles of cv. Hope. Transcript levels of all the six genes were relatively low on day 2, increased from days 3 to 5 and then declined to almost non-detectable levels on day 6. In coleoptiles of CS grown in the light, the early biosynthesis genes (EBGs) were expressed, but the three late biosynthesis genes (LBGs) were not.

Cadmium, copper and zinc ions at high concentrations partially released scarified freshly-harvested seeds ofStylosanthes humilis from physiological dormancy. This response to toxic metals increased along with seed postharvest ageing. Cobalt and silver ions, and abscisic acid impaired metal-promoted germination.

An efficient, highly reproducible system for plant regeneration via somatic embryogenesis was developed for Cenchrus ciliaris genotypes IG-3108 and IG-74. Explants such as seeds, shoot tip segments and immature inflorescences were cultured on Murashige and Skoog (MS) medium supplemented with 2.0-5.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg dm^-3 N⁶-benzyladenine (BA) for induction of callus. Callus could be successfully induced from all the three explants of both the genotypes. But the high frequency of embryogenic callus could be induced only from immature inflorescence explants. Somatic embryos were formed from nodular, hard and compact embryogenic calli when 2,4-D concentration was gradually reduced and BA concentration increased. Histological studies of somatic embryos indicated the presence of shoot apical meristem with leaf primordia. Ultrastructural details of globular and scutellar somatic embryos further validated successful induction and progression of somatic embryogenesis. Shoots were differentiated upon germination of somatic embryos on MS medium containing 2,4-D (0.25 mg dm^-3) and BA or kinetin (1-5 mg dm^-3). Roots were induced on 1/2 MS medium containing charcoal (0.8 %), and the regenerated plants transferred to pots and established in the soil showed normal growth and fertility.

A significant reduction of hop latent viroid (HLVd) content during the generative phase of hop was detected using reverse transcriptase-polymerase chain reaction (RT PCR) and molecular hybridisation methods. A low transmissibility of HLVd through seed may be a feature valuable for the selection and maintenance of viroid-free hybrid hops.

The effects of Cr and Co supplied either individually or mixed together in the nutrient solution on seed germination, enzyme activities, photosynthesis, metabolic products, and yield were investigated. Cr and Co reduced germination percentage only at the highest concentration used but markedly decreased radicle growth which might be attributed to depressive effect of Cr and Co on the activity of amylases and subsequent transport of sugars to the embryo axes. Protease activity, on the other hand, increased with the metal treatment. The highest concentration (10^-2 M) tested of both metals was harmful on plant growth, while the low and moderate concentrations (10^-6 and 10^-4 M) enhanced the contents of chlorophylls and sugars, and activity of Hill reaction. Fresh mass of the produced pods increased at low and moderate concentrations of Cr and at Cr+Co treatment, but decreased in plants treated with Co.

The distribution of the stress-related anionic peroxidase (srPRX) activity was investigated in various organs of cucumber (Cucumis sativus L. cv. Laura) during their development by activity staining and immunoblotting. In shoots, including cotyledon, leaf, stem and tendril, three stress-related peroxidase isoenzymes were present, particularly in old ones. The PRX1 was the only srPRX isoenzyme found in both young and old roots. As fruits became mature, srPRX activity increased dramatically and was particularly enriched in the external parts of the fruit. The PRX1 isoenzyme was highly accumulated in the course of seed germination, while the absence of other two srPRX isoenzymes (PRX2 and PRX3) was recorded. The possible function of the srPRX is discussed, with respect to this spatio-temporal distribution.

Androgenic lines of Brassica juncea cv. PR-45 raised by anther culture, were screen for genetic variation. 393 androgenic plants were transferred to pots to study the R₀ generation. These plants showed substantial variation for different characters. Seed progenies of 27 lines of R₀ plants were sown in the field to study R₁ generation. Androgenic plants within lines were significantly homogeneous for the various agronomic characters studied. Two lines were shorter (18 - 20 %) than the control plants, with a remarkable feature of early maturation. Three lines showed 27 - 31 % higher yield than the parent cultivar.

Two main dormancy states, innate and imposed dormancy, were characterized in turions (winter buds) of the aquatic carnivorous plant Aldrovanda vesiculosa L. (Droseraceae) kept at 3 ± 1 °C in a refrigerator over the winter. As a result of the breaking of imposed dormancy by a temperature increase (at 15 - 20 °C), some of the turions rose to the water surface within 1 - 3 d and germinated. Turion leaves contained large lacunae with a slimy reticulum and were filled by water over winter. As a result of breaking imposed dormancy, the proportion of gas volume in inner turion leaves rose from 10 - 20 % to 100 % of leaf lacunae volume. The aerobic dark respiration rate of the turions [0.74 - 1.5 μmol O₂) kg^-1(FM) s^-1] slightly increased during innate dormancy after 1 - 2 d at 20 °C, while it was almost constant during the breaking of imposed dormancy. The anaerobic fermentation rate of the turions was only 1.5 - 7 % of the oxygen respiration rate and also was constant during the breaking of imposed dormancy. In turions, the content of glucose, fructose, and sucrose was the same for the two states of dormancy, but starch content was greatly reduced for the imposed dormancy (10 - 11 vs. 32 % DM). It may be suggested that a temperature increase causes an increase of fermentation or respiration which is responsible for the evolution of gas in turion lacunae and, thus, for turion rising.

A liquid culture technique has been developed to study lipid metabolism in seeds of Brassica campestris L. grown in vitro from terminal inflorescences detached 4 to 46 days after anthesis. Seeds developed under these conditions exhibited pattern of growth, deposition of storage products and lipid composition similar to those from intact plant.

The physiological function of glutamate dehydrogenase (GDH) was investigated by treating germinating peanut (Arachis hypogaea L.) seeds with nucleoside triphosphate (NTP) solutions in order to alter the isoenzyme distribution patterns. The free nucleosides and nucleotides of the GTP-treated peanut were the highest [8.7 μmol g^-1(f.m.)], and they decreased through the ATP-treated peanut [5.8 μmol g^-1(f.m.)], and CTP-treated peanut [5.5 μmol g^-1(f.m.)], to the UTP-treated peanut [4.1 μmol g^-1(f.m.)]. The combination of 4 NTPs induced 20 % higher content of Pi [173 nmol g^-1(f.m.)] than in the control, but the combined ATP+UTP treatment induced the lowest (93.0 nmol g^-1(f.m.)] Pi. The 4 NTP treatment also induced the highest number of GDH isoenzymes (28) followed by the purine NTP treatments (15 to 20), but the pyrimidine NTP treatments and the combined purine + pyrimidine NTP treatments induced the lowest numbers (<15) of isoenzymes. The deamination/amination ratios were generally higher in the UTP (0.11), and CTP (0.06) treated peanuts than in the GTP (0.04), and ATP (0.07) treated peanuts. There were mutual relationships between higher numbers of GDH isoenzymes present in the GTP-, and ATP-treated peanuts and higher RNA (236.5 and 239.4 μg g^-1, respectively) contents on one hand, and between the lower numbers of isoenzymes in the CTP-, and UTP-treated peanuts and lower RNA (162.0 and 152.5 μg g^-1, respectively) contents. The recurrent relationships of the effects of the NTP treatments of peanut were UTP > ATP > CTP > GTP.

We have assayed different combinations of nutrient media and growth regulators to induce callus and plant regeneration from explants of root, shoot and leaf, complete seed, and isolated mature embryo of barley (Hordeum vulgare L. cv. Hassan). The best results were obtained with mature embryo in J25-8 medium supplemented with 2.0 mg dm^-3 2,4-dichlorophenoxyacetic acid where about 75 % developed friable calli. Some 80 - 85 % of these calli regenerated barley plants in the same J25-8 medium supplemented with 1.0 mg dm^-3 indole-3-butyric acid and 0.1 mg dm^-3 kinetin.

Embryo-rescue was used to facilitate interspecific hybridization of Cucumis anguria L. and C. zeyheri Sond. Embryos were excised from developing fruits at one week intervals for six weeks after hand pollination. Medium containing coconut water was the most suitable for initial germination, and a medium with ascorbic acid was the best for embryo development and plant recovery. Viable plants were obtained from embryos and these plants showed morphological characteristics different from both parents. The analysis of the leucine aminopeptidase (LAP) locus revealed three hybrid types, H1.1, H1.2 and H2.

Heat caused reduction in membrane protein thiol content, increased accumulation of thiobarbituric acid reactive substances and reduced germination rate and early growth in germinating Amaranthus lividus seeds. Imposition of heat stress during early germination also causes accumulation of reactive oxygen species like superoxide and hydrogen peroxide while activities of antioxidative enzymes catalase, ascorbate peroxidase, and glutathione reductase decreased. Calcium chelator (EGTA), calcium channel blocker (LaCl₃) and calmodulin inhibitor (trifluroperazine) aggravated these effects. Added calcium reversed the effect of heat, implying that protection against heat induced oxidative damage and improvement of germination requires calcium and calmodulin during the recovery phase of post-germination events in Amaranthus lividus.

A 51-kDa soluble protein was over-expressed in wheat (Triticum aestivum) seedlings by the treatment of seeds before germination with 50 µM CdCl₂ for 48 h and subsequently washed off Cd²⁺. This protein designated as Cd stress associated protein (CSAP), was purified. Polyclonal antibody was raised against CSAP for localizing the protein in root tissue of treated and control seedlings. It was observed that CSAP was located below the plasma membrane and outer periphery of the tonoplast. This unique type of organized localization of CSAP is suggestive of defensive role against metal phytotoxicity. N-terminal analysis of CSAP and expressed sequence tags (EST) database search of wheat sequences suggests that this protein has not been reported earlier in higher plants.

Efficient plant regeneration through somatic embryogenesis was established for safflower (Carthamus tinctorius L.) cv. NARI-6. Embryogenic calli were induced from 10 to 17-d-old cotyledon and leaf explants from in vitro seedlings. High frequency (94.3 %) embryogenic callus was obtained from cotyledon explants cultured on Murashige and Skoog's germination (MSG) basal medium supplemented with thidiazuron, 2-isopentenyladenine and indole-3-butyric acid. Primary, secondary and cyclic somatic embryos were formed from embryogenic calli in a different media free of plant growth regulators, however, 100 % cyclic somatic embryogenesis was obtained from cotyledon derived embryogenic calli cultured on MSG. Somatic embryos matured and germinated in quarter-strength MSG medium supplemented with gibberellic acid. Cotyledons with root poles or non root poles were converted to normal plantlets and produced adventitious roots in rooting medium. Rooted plants were acclimatized and successfully transferred to the field.

Inflorescence explants of two winter wheat cultivars, Triticum durum cv. Kiziltan-91 and T. aestivum cv. Bezostaja-01, were used to evaluate the effects of vernalization period of donor plants, callus age and medium composition on regeneration capacity. Donor plants were grown for 7 d and they were exposed to 4 °C for 1, 2, 3, 4, and 5 weeks. The maximum inflorescence formation was observed as 79 % at 4 weeks and 73 % at 5 weeks of vernalization period for Kiziltan-91 and Bezostaja-01, respectively. Among 6 different callus induction and regeneration mediums, I₁-R₁ and I₃-R₃ have to be the best responding mediums for Kiziltan-91 and Bezostaja-01, respectively. In Kiziltan-91, calli induced from donor plants, vernalized for 3 weeks, showed a significantly lower regeneration capacity than counterparts vernalized for 4 and 5 weeks. The highest regeneration capacity of 69 % was obtained from 6-week-old calli produced from 4 weeks vernalized Kiziltan-91 donor plants. In contrast to Kiziltan-91 cultures, the effects of vernalization period and callus age on regeneration capacity were not significant in Bezostaja-01 cultures. The maximum numbers of tillers were obtained from 6-and 15-week-old calli for Bezostaja-01 and Kiziltan-91, respectively. In contrast to vernalization period of donor plants, callus age had no effect on seed number.

Using different explants of in vitro seed grown Scutellaria baicalensis Georgi plantlets, hairy roots were induced following inoculation of Agrobacterium rhizogenes strains A₄GUS, R1000 LBA 9402 and ATCC11325. The A₄GUS proved to be more competent than other strains and the highest transformation rates were observed in cotyledonary leaf explant (42.6 %). The transformed roots appeared after 15-20 d of incubation on hormone free Murashige and Skoog medium. Growth of hairy roots was assessed on the basis of total root elongation, lateral root density and biomass accumulation. Maximum growth rate was recorded in root:medium ratio 1:100 (m/v). Hairy root lines were further established in Gamborg B₅ medium and the biomass increase was maximum from 15 to 30 d. PCR, Southern hybridization and RT-PCR confirmed integration and expression of left and right termini-linked Ri T-DNA fragment of the Ri plasmid from A₄GUS into the genome of Scutellaria baicalensis hairy roots. GUS assay was also performed for further integration and expression. All the clones showed higher growth rate them non-transformed root and accumulated considerable amounts of the root-specific flavonoids. Baicalin content was 14.1-30.0 % of dry root mass which was significantly higher then that of control field grown roots (18 %). The wogonin content varies from 0.08 to 0.18 % among the hairy root clones which was also higher than in non-transformed roots (0.07 %).

A micropropagation protocol through multiple shoot formation was developed for Thlaspi caerulescens L., one of the most important heavy metals hyperaccumulator plants. In vitro seed-derived young seedlings were used for the initiation of multiple shoots on Murashige and Skoog (MS) medium with combinations of benzylaminopurine (BA; 0.5-1.0 mg dm^-3), naphthaleneacetic acid (NAA; 0-0.2 mg dm^-3), gibberellic acid (GA₃; 0-1.0 mg dm^-3) and riboflavin (0-3.0 mg dm^-3). The maximum number of shoots was developed on medium containing 1.0 mg dm^-3 BA and 0.2 mg dm^-3 NAA. GA₃ (0.5 mg dm^-3) in combination with BA significantly increased shoot length. In view of shoot numbers, shoot length and further rooting rate, the best combination was 1.0 mg dm^-3 BA + 0.5 mg dm^-3 GA₃ + 1.0 mg dm^-3 riboflavin. Well-developed shoots (35-50 mm) were successfully rooted at approximately 95 % on MS medium containing 20 g dm^-3 sucrose, 8 g dm^-3 agar and 1.0 mg dm^-3 indolebutyric acid. Almost all in vitro plantlets survived when transferred to pots.

A protocol for multiple shoot induction from cotyledonary node explants of Terminalia chebula Retz. has been developed. Germination frequency of embryos (up to 100 %) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg dm^-3 gibberellic acid (GA₃). Maximum number of shoots (6.4 shoots per cotyledonary node) was obtained on half-strength MS + 0.3 mg dm^-3 GA₃+ 1.0 mg dm^-3 indole-3-butyric acid (IBA) + 10.0 mg dm^-3 benzylaminopurine (BAP) after 4 weeks of culture. When the cotyledonary nodes along with the axillary shoot buds were allowed to grow in the same medium upto 19.2 shoots were obtained after 8 - 9 weeks. Best rooting (100 %, 5.5 roots per shoot) was observed when shoots were excised and transferred to half-strength MS medium containing 1.0 mg dm^-3 IBA + 1 % mannitol and 1.5 % sucrose. Survival of rooted plants in vivo was low (35 - 40 %) when they were directly transferred to soil in glasshouse. However, transfer to soil with MS nutrients and 1.0 mg dm^-3 IBA in culture room for a minimum duration of 2 weeks increased the survival percentage of plants to 100 %.

Thirteen flavonoid aglycons, contained in the strongly allelopathic epicuticular exudates of Dittrichia viscosa, were investigated for their effects on lettuce seedling radicle growth. Concerning radicle length and mass, variable results were obtained, with most of the substances having no effect, some being inhibitory and some even promotive. Shoot mass was slightly reduced in four cases. Seed germination rates, root hair and lateral root formation were not affected either. Three of the compounds (namely quercetin 3,3-dimethylether, naringenin and eriodictyol) induced a strong ageotropic response in radicle growth.

Heat shock (HS) reduced total lipid and phospholipid contents and their synthesis in germinating seeds of pigeonpea [Cajanus cajan (L.) Millspaugh]. Lipid peroxidation was also enhanced with increasing temperature and HS duration. HS influenced lipid metabolism to a higher extent at 45°C than at 40°C. This altered lipid metabolism and lipid peroxidation was associated with the loss of various solutes from the germinating seeds, and modification of growth and development. Pretreatment of germinating seeds at 40°C for 1 h or at 45°C for 10 min followed by incubation at 28°C for 3 h prior to 45°C for 2 h ameliorated solute leakage due to reduced lipid peroxidation and improvement in lipid content and membrane function.

Seed storage proteins of Ebenus cretica were fractionated to albumins, globulins, prolamins and glutelins according to their solubility in water, 0.5 M NaCl solution, 55 % propanol-2 and 0.125 M sodium borate (pH 9.0) containing 0.5 % SDS (sodium dodecyl sulfate) solution, respectively. Glutelins consist of the major (about 81 %) fraction of the total extracted proteins. Analysis by SDS-PAGE revealed that the total extracted protein patterns from different racemes of the same plant were similar, while those from seeds of different plants were different. In addition, distinct differences were observed within protein patterns of alkaline extractable glutelin fractions and salt soluble globulin fractions. In E. cretica four ecotypes (A - D) were distinguished by SDS-PAGE of total extracted seed proteins. The last method was more simple and rapid than others and was suggested for screening analysis.

Wheat (Triticum aestivum L.) plants were subjected to mild water stress during grain filling at milk (early, medium, and late) and dough (early, soft, hard) stages. The grains harvested from stressed plants were subjected to low temperature stress of 10 °C for 24 h in presence or absence of 1 mM CaCl₂, and embryos were examined for oxidative injury. The embryos of grains water stressed at milk and soft dough stages showed lowest contents of H₂O₂ and malondialdehyde and highest membrane stability index, ascorbic acid content, and activities of catalase, ascorbate peroxidase, and superoxide dismutase as compared to control embryos or water-stressed at other stages. Presence of Ca²⁺ in the medium reduced H₂O₂ and malondialdehyde content and increased ascorbic acid content, and catalase, ascorbate peroxidase and superoxide dismutase activities.