Combined Effect of Some Growth Regulators on Growth and Gametangial Formation in the Liverwort Riccia gangetica Ahmad

The present work deals with the effect of IAA-kinetin, IAA-GA³and GA³-ABA interactions on growth and gametangial formationin Riccia gangetica in vitro. Inhibitory effect of high concentrationof IAA on vegetative growth is overcome by the co-addition ofkinetin. The best response in terms of fresh and dry weightyields of thalli is obtained by a combination of 10^–5mol dm^–3 kinetin+ 10^–7 mol dm^–3 IAA. Interactionof IAA and kinetin has an additive effect on archegonial formation.Co-addition of IAA and GA³ decreases production of archegoniaand antheridia as compared to those produced in response toIAA and GA³ alone, respectively. Combination of GA³ and ABAreduces vegetative growth, as well as the number of anthendiaand archegonia. Key words: Riccia gangetica, growth, growth regulators, gametangial formation     

MORPHOGENESIS IN SPIEODELA OLIGORRHIZA: EFFECTS OF PYRIMIDINE BASE ANALOGUES ON INITIATION, ELONGATION AND DIFFERENTIATION OF FRONDS

The effect of four pyrimidine base analogues and their antidoteson S. oligorrhiza was studied. FUdR stopped cell division at concentrations of 4 10^–7M and higher. This effect could be nullified by the additionof 4 10^–6 M thymidine. Neither uridine nor uracil hadan antidotal effect on FUdR. FU (8 10^–6 M or higher concentration) affected celldivision, frond elongation and differentiation, and could notbe counteracted by either thymidine or uracil. TU (8 10^–4 M) rather specifically inhibited differentiationof frond tissues, while not preventing cell division or theinitiation of new generations. Uracil and uridine at about equimolarconcentrations completely counteracted the TU effect. AzU (10^–3 M) suppressed cell division, frond elongationand frond differentiation. When thymidine (10^–3 M) wasadded simultaneously with AzU only cell elongation and differentiationof fronds were inhibited, but cell division proceeded. 10^–3M uracil (but not uridine) counteracted all effects of AzU. ¹ Based on a portion of the senior author's Ph.D. Thesis.     

In vitro Propagation of Red Cabbage (Brassica oleracea L. var. capitata)

By manipulation of various growth regulators and physical conditions,plants have been regenerated from excised roots, stem segments,cotyledons, leaves, and callus cultures of red cabbage (Brassicaoleracea var. capitata) grown under in vitro conditions. Shootbuds were induced on isolated root segments (1 cm long) culturedon Murashige and Skoog's medium and the frequency of bud formationwas greatly enhanced by the addition of kinetin (0.5 part 10^–6).Callus obtained from the seeds, cotyledons, and hypocotyl segmentscultured on a medium fortified with 2,4-D (1 part 10^–6),kinetin (0.1 part 10^–6), and coconut milk (10%, v/v) hasbeen repeatedly subcultured. The callus is slow growing, andon transference to a kinetin (2 parts 10^–6) and IAA (2parts 10^–6) medium underwent morphogenesis to give riseto plants. The significance of the propagation of red cabbageby in vitro culture is pointed out.     

Effect of abscisic acid on elongation and kinetin-induced tuberization of isolated stolons of Solarium tuberosum L.

The response of isolated stolons cultured in vitro, to abscisicacid (ABA) has been studied in the presence and absence of kinetin(6-furfurylaminopurine). ABA alone in concentrations from 7.5x 10^–4 mM to 7.5 x 10^–2 mM, inhibited stolon elongationbut failed to promote tuber initiation. In the presence of kinetin,ABA at concentrations of 3.0 x 10^–2 and 7.5 x 10^–2mM markedly inhibited kinetin-induced tuber initiation and stolonelongation, but at 7.5 x 10^–4 and 7.5 x 10^–3 mMABA did not prevent tuber initiation. When stolons were incubated on a medium containing kinetin andlater transferred to one containing ABA with or without kinetin,the inhibitory effect of ABA decreased appreciably as the timeof incubation on kinetin is increased. The results are discussed in relation to the role of ABA inthe inhibition of nucleic acid and protein synthesis and theinteraction with cytokinins and the possible effect of ABA onkinetin uptake, transport and accumulation at the locus of action. (Received February 26, 1969; )     

Effect of External Supply of Growth Substances on Axillary Proliferation and Development in Dioscorea bulbifera

Bulbil development in cultured nodes of D. bulbifera proceededin the absence of growth substances from the medium. When IAAwas incorporated into the medium at the concentrations of 5mg l^–1 and 10 mg l^–1 the cultured nodes producedlarger bulbils than in its absences. When the concentrationof IAA was increased to 15 mg l^–1, however, the culturednodes produced a callus instead of a properly organized bulbil.The dry weight of bulbils increased when kinetin was added tothe medium at the concentrations of 0.05, 0.5, and 2.5 mg l^–1.The greatest increase was with 0.5 mg l^–1 kinetin. Onincreasing the concentration of kinetin in the medium to 5.0mg l^–1 the tissue produced had smaller dry weight thanthose produced in the absence of growth substances. Additionof different combinations of IAA and kinetin to the basal mediumresulted in the production of normal bulbils, roots, and shootsin some instances (suitable combinations) and in the productionof callus and abnormal shoots in others (non suitable combinations).     

Auxin-gibberellin relationships in their effects on hypocotyl elongation of light-grown cucumber seedlings VI. Promotive and inhibitory effects of kinetin

The interaction of kinetin with IAA and GA₃ on the elongationof hypocotyl sections of Cucumis sativus L. cv. National Picklingwas studied. Kinetin in the concentration range of 10^–7M to 10^–4 M markedly inhibited IAA-induced elongation,while in a lower range from 10^–10 M to 10^–8 M, itsynergistically enhanced IAA-induced elongation. Kinetin alonein this range had no effect. A 5-to 15-min pulse treatment seemsenough to induce the maximum effect for both inhibition andpromotion. Since the magnitude of the maximum inhibition dependedon the concentration and not on the duration of treatment, thereaction in the cell caused by kinetin seemed to be completedwithin a short period. Washing of the sections with distilledwater after kinetin treatment (30 min) did not significantlyeliminate the kinetin effect. This probably indicates that thebinding of kinetin molecules to a supposed acceptor is not reversible.Interaction of kinetin with GA₃ in their pretreatment effectson IAA-induced elongation shows that in the inhibitory concentrationrange, the kinetin effect was partly overcome by GA₃, and thatin the promotive range, the magnitude of the enhancement wasdetermined by kinetin regardless of the presence of GA₃. Theeffect of kinetin seems to dominate over that of GA₃ indicatingthat the modes of their pretreatment effects differ from oneanother. (Received June 24, 1977; )     

Involvement of cellulose synthesis in actions of gibberellin and kinetin on cell expansion. Gibberellin-coumarin and kinetin-coumarin interactions on stem elongation

In azuki bean (Azukia angularis = Vignia angularis) epicotylsections, 5 ? 10^–4 M coumarin inhibited the incorporationof radioactivity from [U^–14C]glucose into the cellulosefraction by 35% in the absence of indole-3-acetic acid (IAA)and by 40% in the presence of 1 ? 10^–4 M IAA. There wasno inhibitory effect on the incorporation of radioactivity intothe other fractions. Coumarin at 5 ? 10^–4 M reversed thepromoting effect of 1 ? 10^–5 M gibberellin A₃ (GA) andthe inhibitory effect of 1 ? 10^–5 M kinetin on IAA-inducedelongation of sections with no significant effects on IAA-inducedelongation. Neither GA nor kinetin had any appreciable effectson cellulose synthesis. No inhibition of cellulose syntheiswas observed with 1 ? 10^–3 M colchichine, which has beenreported to have effects similar to those of coumarin on GA-or kinetin-affected stem elongation. Coumarin at 5 ? 10^–4M was ineffectual in breaking up wall microtubules, while adisrupting effect on wall microtubules was clearly demonstratedwith 3 ? 10^–4M colchicine. From these results, the possible involvement of cellulose synthesisin cell expansion controlled by GA or kinetin was suggested. (Received August 3, 1973; )     

Hormonal Control of Xylogenesis in Pith Parenchyma Explants of Lactuca

The time course of xylem differentiation was determined in explantsof lettuce pith parenchyma (Lactuca sativa L. cv. Romana) culturedon Murashige and Skoog (1962) medium using different concentrationsof auxin (IAA) and one cytokinin (zeatin or kinetin). Increasinglevels of auxin from I mg 1^–1 to 15 mg 1^–1 in thepresence of a constant level of a cytokinin (zeatin or kinetin)yielded up to 10 mg 1^–1 IAA, an increase in the numberof tracheary element formations. Cytokinin concentrations aboveand below o.1 mg 1^–1 interacting with an optimal xylogenicamount of auxin inhibited xylogenesis. The IAA (10 mg 1^–1)-zeatin(0.1 mg ^1–1) treatment produced the greatest number oftracheids, while kinetin compared to zeatin did not producesuch an effect. The different effectiveness of zeatin and kinetinin inducing tracheary element formations was not due to a differentcapacity of the two cytokinins to stimulate cell division butit seems likely that zeatin, because of interaction with IAA,is more active than kinetin in the determination of the dividingcells in a specific type of cytodifferentiation. The IAA (10mg 1^–1)-zeatin (0.1 mg 1^–1) treatment produced about6.9 per cent tracheids with respect to cell division while IAA(10 mg 1^–1)-kinetin (0.1 mg 1^–1) produced 4.2 percent. These results are discussed with reference to the problemsof hormonal control of xylem differentiation.     

Effects of Kinetin on Growth of the Apical Meristem and Floral Differentiation in Chenopodium rubrum L.

Kinetin (1•10^–4 M and 1•10^–3 M) was appliedto the plumules of 6-day-old Chenopodium rubrum plants. Effectson growth, anatomical structure and organogenesis in the apicalmeristem were followed. Floral differentiation as affected bykinetin was also investigated in plants induced to flower byshort-day treatment. Kinetin increased mitotic activity in the apical meristems inboth induced and non-induced plants. The effect was most pronouncedin the peripheral and subcentral zone. An increase in nucleolussize and a higher degree of pyroninophilia in the peripheralzone was also observed, indicating a localized promotion ofRNA synthesis. A higher rate of leaf initiation and a stimulationof leaf and stem growth was subse quentiy recorded. The growthof axillary meristems and of bud primordia was promoted onlyat the lower concentration of kinetin (1•^10–4 M),in both photoperiodically-induced and non-induced plants. However,the pattern of lateral bud growth differed from that found innormal floral differentiation. In kinetintreated plants, thebud primordia are isolated from the summit of the shoot apexby a succession of rapidly growing leaves. The enhancement ofleaf growth leads to correlative inhibition of axillary budpriniordia and results, finally, in a suppression of floraldifferentiation. The inhibitory effect of kinetin on floweringwas compared with that of auxin. Inhibition of flowering occurredin both cases but is achieved in two different ways.     

Differentiation of Shoot Buds in vitro in Tissue 1Sisymbrium irio L.

Shoot bud formation was induced in the stem callus of Sisymbriumirio L., a Cruciferous plant. The callus was established onMurashige and Skoog medium with IAA (1?0 mg l^–1) and kinetin(0?5 mg l^–1). The effect of three purines (kinetin, 6-benzylaminopurine,and 6-methylaminopurine) incorporated singly along with IAAin MS medium was investigated. It was found that kinetin orMAP (3–5 mg l^–1) along with IAA (0?5 mg l^–1)were the most effective in inducing shoot bud formation. Adeninesulphate (10 mg l^–1) with kinetin (1?0 mg l^–1) alsoinduced bud differentiation. The morphogenetic potential of the callus to differentiate shootbuds was seemingly lost in 2 year old callus cultures. However,on successively subculturing on a regeneration medium shootbuds differentiated and the number of buds formed improved onfurther subculture. Two types of meristematic outgrowths were recognized: (i) arisingfrom superficial cells and (ii) arising from deep-seated cellsin the vicinity of tracheidal elements. However, both typesformed meristematic nodules on the surface of which shoot budsdifferentiated. Some embryoids were also recognized arisingsuperficially.     

Investigation of the Radish Leaf Bioassay for Kinetins, and Demonstration of Kinetin-like Substances in Algae

This study is part of an investigation into the occurrence ofplant growth substances in marine unicellular algae. Auxinsand gibberellins have previously been detected. This paper reportsthe occurrence of phytokinins in algae, using the radish leaftest. The technique of the test is described and a few modificationsexamined. Radish leaves were shown to be more responsive thanswede leaves. Some requirement for a minimum temperature below7? C during the growing period of the plants appeared to bea factor for maximum response. The size and age of the leafwere shown to influence kinetin activity. Chemicals tested includedbenzyl adenine, a 9-substituted benzyl adenine, and kinetin.Kinetin showed lower activity than the benzyl adenines. Kinetinkept in the solid state at –20? C for 2 years was as activeas freshly prepared kinetin. Gibberellic acid (GA₂) at 10^–5g/ml sometimes showed activity equivalent to 10^–5 g/mlkinetin. Indole-3-yl acetic acid at concentrations of 10^–4to 10^–6 g/ml was inactive. The radish leaf test was successfully used to demonstrate phytokininsin extracts of two species of unicellular marine algae, andin marine phytoplankton samples. Amounts found were within therange 0.1 to 1.0 mg/kg with one exception of 10 mg/kg. Phytokininactivity in these extracts decreased over a period of a fewweeks when stored at –20? C.     

Cyclic Nucleotides and In Vitro Plant Cultures: I. INDUCTION OF ORGANOGENESIS IN TOBACCO (NICOTIANA TABACUM CALLUS CULTURES

Mangat, B. S. and Janjua, S. 1987. Cyclic nucleotides and invitro plant cultures. I. Induction of organogenesis in tobacco(Nicotiana tabacum) callus cultures.—J. exp. Bot. 38:2059–2067. The possibility that cyclic nucleotides have a mediatory rolesimilar to cytokinins in plant tissue cultures was examined.Calli obtained from tobacco pith tissue were incubated on growthmedia supplemented with either cyclic AMP, cyclic GMP, adenosineor guanosine, in concentrations ranging from (mg dm^–3)0 to 2·0 together with 2·0 mg dm^–3 of IAA.Results were compared with identical calli grown on media containingcomparable amounts of kinetin and IAA. Increase in callus growthwas observed on all media containing cyclic AMP, cyclic GMP,adenosine, guanosine or kinetin. Adenosine or guanosine didnot promote organogenesis. Low concentrations (0·02 and0·05 mg dm^–3) of kinetin stimulated extensive rootdevelopment. Some root formation was also elicited with higheramounts of cyclic AMP (0·1 and 0·2 mg dm^–3)or cyclic GMP (0·2 and 0·5 mg dm^–3). Bothkinetin and cyclic GMP promoted shoot differentiation. However,in contrast to kinetin, cyclic GMP induced organogenesis atlower concentrations (0·02 and 0·1 mg dm^–3).The addition of 2·0 mg dm ^–3 of cyclic AM P toIAA-free growth media elicited shoot differentiation. This wasalso the case with a similar concentration of kinetin or cyclicGMP. Results suggest cytokinin activity for the two cyclic nucleotides. Key words: Tobacco, Nicotiana tabacum, tissue culture, cyclic nucleotides, cyclic AMP, cyclic GMP organogenesis     

The Effects of Cytokinins and ABA on Stomatal Behaviour of Maize and Commelina

Epidermal strips and leaf fragments of Commelina and leaf fragmentsof maize were incubated on solutions containing naturally-occurringor synthetic cytokinins and/or ABA. The effects of these treatmentson stomatal behaviour were assessed. Cytokinins alone did notpromote stomatal opening in either species but concentrationsof both zeatin and kinetin from 10^–3 to 10^–1 molm^–3 caused some reversal of ABA-stimulated closure ofmaize stomata. The reversal of the ABA effect increased withincreasing cytokinin concentration. Cytokinins had no effecton ABA-stimulated closure of Commelina stomata. When appliedalone, at high concentration (10^–1 mol m^–3), toCommelina epidermis or leaf pieces both zeatin and kinetin restrictedstomatal opening. Key words: ABA, Cytokinins, Stomata, Maize, Commelina     

Effect of Some Known Growth Regulators on Growth and Fertility in Male Clones of the Moss Microdus brasiliensis (Dub.) Ther.

Among the auxins (IAA, 2,4-D, NAA and NOA) LAA proves inhibitoryfor antheridial formation. The rest promote this response, andNAA is most effective. Cytokinin (2iP) stimulates vegetativegrowth as well as antheridial formation, but the effect is morepronounced on the former. Interaction of kinetin and IAA provesbetter for antheridial production as compared to IAA alone.Gibberellic acid enhances gametangial induction as well as vegetativegrowth at lower levels (10^–8–10^–8 mol dm^–3).With abscisic acid both the responses are markedly reduced.Anti-auxins and cycocel promote antheridial production and vegetativegrowth. Testosterone is more potent than progesterone in promotingantheridial formation and vegetative growth. Key words: Fertility, growth hormones, moss     

Effect of Plant Growth Regulators on Nitrate Utilization by Roots of Nitrogen-Depleted Dwarf Bean

We examined the effect of pretreatments (18 h at 5 µmoldm^–3) with abscisic acid, the ethylene-releasing substance‘Ethephon’, gibberellic acid, indoleacetic acid,kinetin and zeatin on nitrate uptake and in vivo nitrate reductaseactivity (NRA) in roots of nitrogen-depleted Phaseolus vulgarisL. Nitrate uptake showed an apparent induction pattern witha steady state after about 6 h, in all treatments. The nitrateuptake rate after 6 h was unaffected or at most 30% lower aftertreatments with the plant growth regulators. Gibberellic acid, kinetin and zeatin induced substantial NRAin roots in the absence of nitrate, whereas Ethephon enhancedNRA only during nitrate nutrition. Kinetin-induced NRA (Ki-NRA)was maximal after a pretreatment at 1 µmol dm^–3,and showed a lag phase of 6–8 h. Ki-NRA was additive tonitrate-induced NRA (NO^–₃-NRA) for at least 24 h, independentof the induction sequence. After full induction, Ki-NRA approximated20% of NO-₃-NRA. Abscisic acid counteracted the developmentof Ki-NRA, but not of NO^–₃-NRA. Cycloheximide and tungstatewere equally effective to suppress the development of nitratereductase activity after supply of kinetin or NO^–₃. Our data are consistent with the operation of two independentenzyme fractions (Ki-NRA and NO^–₃-NRA) with apparentlyidentical properties but with separate control mechanisms. Theabsence of major effects of plant growth regulators on the time-courseand rate of nitrate uptake suggests that exogenous regulators,and possibly endogenous phytohormones are of minor importancefor initial nitrate uptake. The differential effect of someregulators on nitrate uptake and root NRA furthermore indicatesthat the processes of uptake and reduction of NO^–₃ arenot obligatory or exclusively coupled to each other.     

EFFECTS OF CHLORAMPHENICOL AND KINETIN ON UPTAKE AND INCORPORATION OF AMINO ACIDS BY TOBACCO LEAF DISKS

The effects of chloramphenicol and kinetin on uptake and incorporationof ³⁵S-methionine and some ₁₄C-amino acids have been investigatedin leaf-disks of Nicotiana rustica in light and dark. Chloramphenicolin a concentration of 1 mg per ml inhibits the uptake of aminoacids from 30 to 60 per cent compared with the water control.The incorporation of amino acids into bulk protein is stronglyinhibited in light (40 to 70 per cent), but only to a smalldegree in dark (10 to 20 per cent), as revealed also by ¹⁴CO₂-photosynthesisof the disks and following treatment with chloramphenicol indark. The stimulating effect of kinetin on uptake and incorporationof amino acids is dependent upon its concentration (10^–5to 10^–6 M ; but 10^–4 M solution inhibits stronglyboth uptake and incorporation). The stimulation seems to influencemore incorporation than uptake processes. Possible interactionsof chloramphenicol and kinetin in the protein metabolism oftobacco leaves have been discussed. (Received April 27, 1964; )     

Isolation, Culture, and Plant Regeneration of Protoplasts of Two Shrubby Oxalis Species from South America

Protoplasts were successfully isolated from internodal callustissues of both Oxalis glaucifolia and O. rhombeo-ovata whenthey were digested in a solution containing 0.1% (w/v) MacerozymeR-10, 0.5% (w/v) cellulase Onozuka R-10 and 0.3 mmol m^–3sucrose. Protoplasts proliferated to give cell colonies on Gamborget al.'s B5 medium supplemented with 0.3 mmol m^–3 mannitol,0.5 mg dm^–32, 4-D, and 2.0 mg dm^–3 kinetin. Calluswas produced upon transfer of cell colonies to Murashige andSkoog medium containing 2.0 mg dm^–3 l-naphthaleneaceticacid (NAA) and 0.1 mg dm^–3 kinetin for O. glaucifolia,or with 5.0 mg dm^–3 NAA and 0.5 mg dm^–3 6-benzylaminopurine,for O. rhombeo-ovata. Plants were regenerated from O. glaucifoliaprotoplasts on a medium containing 0.1 mg dm–3 NAA, 1.0mg dm^–3 kinetin and 1.0 mg dm^–3 gibberellic acid,but only vascular nodules were differentiated by O. rhombeo-ovataprotoplast-derived calli. Key words: Tissue culture, protoplasts, plant regeneration, Oxalis spp     

The Effect of Salinity on Germination and Its Correlation with K+, Na+, Cl- Accumulation in Germinating Seeds of Triticum aestivum L. cv. Akbar

NaCl salinity stress consistantly decreased the rate of germinationof wheat. GA alone or in combination with kinetin alleviatedthe inhibitory effect of salinity on germination. However, kinetinfurther decreased the rate of germination under NaCl salinitystress. NaCl salinity increased accumulation of Na⁺ and Cl^–while it decreased K⁺ accumulation in germinating seeds. GAcaused an increase in K⁺ accumulation and a decrease in Cl^–accumulation in the germinating seeds while kinetin increasedCl^– accumulation in salinity stressed plants. The co-relationbetween the effect of salinity on germination and that on accumulationof ions is discussed. (Received February 12, 1992; Accepted August 4, 1992)     

CHANGES IN RESPONSE TO PURINE AND PYRIMIDINE DERIVATIVES OF CARROT ROOT CALLUS DURING SUCCESSIVE CULTURING

1. The growth of the carrot root callus which had been subculturedfor a long period (CCL) was promoted by the addition of 5l0^–8and 5l0^–7 M kinetin, whereas in the callus subculturedfor a short period (CCS) no growth promotion was observed atany concentrations of kinetin tested.
2. CCL showed an increasedgrowth in response to the applicationof kinetin, guanine, adenine,hypoxanthine, uracil, thymine,and cytosine in the presenceof fractions A and C of carrotroot extract, whereas no suchresponse was observed in CCS.CCL required fraction C to respondto uracil and probably purineand pyrimidine derivatives ingeneral.
3. The growth of CCL was promoted by kinetin, guanine,adenine,or hypoxanthine in the medium containing inositol andaminoacids mixture. In this case the growth-promoting actionof guanine,adenine, or hypoxanthine was nullified by kinetin.

(Received December 24, 1964; )