Anwar A. Khan
ACC-derived ethylene production was used as an index of seed vigor of lettuce (Lactuca sativa L.), cabbage [Brassica oleracea (Capitata Group)], tomato (Lycopersicon esculentum Mill.), snap bean (Phaseolus vulgaris L.), and sweet corn (Zea mays L.) seeds. Seeds were aged at 40C and 93% relative humidity over saturated solution of KH2PO4 for various times to obtain seeds of differing vigor. Naturally aged lettuce seeds, differing in vigor, were also used. Depending on the seed type, 0.25 to 2 mm ACC (saturating dose) was needed to produce maximal amounts of ethylene. Seeds in the presence of ACC produced a much larger amount of ethylene than those in the absence of ACC, the ACC-derived ethylene could be detected before germination, and ACC had no adverse effect on germination. ACC-derived ethylene production paralleled vigor loss as determined by a decrease in percentage germination over a soak period required for complete germination of nonaged seeds (16 hours for lettuce, 24 hours for cabbage, and 48 hours for tomato and sweet corn), an increase in mean germination time (determined for lettuce only), and a decrease in seedling growth (determined for snap bean only). Second degree polynomial and logarithmic equations generated for the relationship of ACC-derived ethylene production to germination or growth parameters following seed aging, provided good to excellent fit. As a vigor test, the ACC-ethylene procedure has several advantages over the non-ACC ethylene procedure: It improves the sensitivity of the test by enhancing ethylene production, permits detection of small differences in vigor, and allows detection of ethylene before germination within a few hours of soaking. Chemical name used: 1-aminocyclopropane-1-carboxylic acid (ACC).
Anwar A. Khan
Quantification of seed dormancy has been achieved by measuring physiological, biochemical, and molecular changes accompanying dormancy release, as well as dormancy development. At the physiological level, dormancy is quantified in terms of stratification time, strength of embryo covering structures, embryo growth potential, responsiveness to light, and to temperatures and other changes. At the biochemical level, dormancy has been related to hormone (abscisic acid, gibberellin, etc.) levels, respiratory activity, and other metabolic functions. At the molecular and cellular level, dormancy has been associated with RNA and protein synthesizing ability and with gene expression. Our recent studies with lettuce seeds using gibberellin biosynthesis inhibitors indicate that the amount of gibberellin produced during seed soak may mediate dormancy release and is quantitatively related to the level of dormancy. Examples of quantifiable changes associated with dormancy will be described. Whether a quantifiable change reflects a causal relationship with dormancy release or development, or is a consequence thereof, will be discussed.
Anwar A Khan
Fluridone (FL), a catotenoid biosynthesis inhibitory herbicide, prevented supraoptimal temperatures from inducing thermodormancy in seeds and permitted seedling emergence in several lettuce cultivars. A 48-h dark soak at 35C with 23 μM FL completely prevented the induction of thermodormancy in `Mesa 659' and `Emperor' lettuce seeds as more than 90% of the seeds germinated on transfer to water in darkness at 25C. Abscisic acid (100 μM) applied with FL did not prevent FL from acting. Dormancy was released completely in the naturally dormant `Garnet' and `Grand Rapids' lettuce seeds at 25C in darkness by 1 μM FL. FL applied following thermodormancy induction at 35C in `Mesa 659' lettuce seeds had little effect on releasing the induced dormancy. A 2-h presoak of `Mesa 659' lettuce seeds with 0.47 μM FL followed by washing, drying, and sowing in a peatlite mix at 25C/35C (12 h/12 h), permitted 80% seedling emergence. Higher concentrations resulted in the emergence of albino seedlings. Similar results were obtained with seeds of other lettuce cultivars (`Prizehead', `Emperor', `Ithaca', and `Empress'). A 6-h matriconditioning (MC) (A.A. Khan, Hort. Rev. 13:131–181, 92) of `Mesa 659' lettuce seeds in 7.5 μM FL enhanced the emergence to a greater extent than by MC alone at 25C/35C. The FL procedure in alleviating high-temperature stress in lettuce and other seeds is being tested further.
Anwar A. Khan
A gibberellic acid (GA) biosynthesis inhibitor, tetcyclacis, induced dormancy in nondormant seeds of lettuce (Lactuca sativa L.), tomato (Lycopersicon esculentum Mill.), pepper (Capsicum annuum L.), carrot [Daucus carota var. sativus (Hoffn.)], onion (Allium cepa L.), celery (Apium graveolens L.), and impatiens (Impatiens novette), as most of the seeds failed to germinate after washing under conditions that permitted germination before dormancy induction. In lettuce seeds, tetcyclacis and paclobutrazol were more effective in inhibiting germination in light than in darkness. A 16- to 24-h soak treatment with tetcyclacis was sufficient to induce dormancy in nearly all seeds. Tetcyclacis failed to induce dormancy if applied after 6 h presoak in water. Dormancy induced by tetcyclacis was released by GA4+7 (a mixture of gibberellin A4 and A7), light, and moist-chilling treatments. When GA4+7 was applied with tetcyclacis, dormancy induction was prevented under both favorable, e.g., 25C, and unfavorable, e.g., 5C, or low water potential (Ψ), germination conditions. Unlike tetcyclacis, abscisic acid (ABA) failed to induce dormancy in lettuce seeds. Thermodormancy induction in lettuce seeds at 35C was prevented by fluridone. However, neither ABA nor tetcyclacis countered its effect. Dormancy was also induced in lettuce seeds by ancymidol, flurprimidol, or paclobutrazol. Dormancy induced by tetcyclacis in pepper, tomato, carrot, and onion seeds was released by GA4+7, but not by irradiation or moist-chilling. Chemical names used: 5-(4-chlorophenyl)-3, 4, 5, 9, 10-pentaazatetracyclo [5.4.102,6.08,11]-dodeca-3, 9-diene (tetcyclacis); 1-(4-chlorophenyl)-4, 4-dimethyl-2-(1H-1, 2, 4-triazole-1-yl)-3-pentanol (paclobutrazol); α-cyclopropyl-α-(4-methoxyphenyl)-5-pyrimidine methanol (ancymidol); α-(1-methyl)-α-[4-(trifluoromethoxy) phenyl]-5-pyrimidine-methanol (flurprimidol); 1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4 (1H)-pyridinone (fluridone).
Anwar A. Khan
1-aminocyclopropane-1-carboxylic acid (ACC) was used to measure seed vigor of lettuce, cabbage, tomato, snap bean and sweet corn seeds. Accelerated aging at 40C and 93% RH over saturated solution of KH2PO4 and natural aging under ambient storage conditions (5-7C, 28 to 60% RH) were used to obtain seeds of differing vigor levels. Depending on the type of seeds, the amount of ACC needed for maximal ethylene production (saturating dose) ranged from 0.25 to 2mM. Seeds produced much larger amounts of ethylene in the presence than in absence of ACC, the ACC-derived ethylene was detected prior to germination, and ACC had no adverse effect on germination. ACC-derived ethylene production paralleled vigor loss as determined by percentage germination, mean germination time (for lettuce only) and seedling growth (for snap bean only). Second degree polynomial and logarithmic equations generated for the relationship of ACC-derived ethylene production to germination or growth parameters following seed aging, provided good to excellent fit. As a vigor test, the ACC-ethylene procedure has several advantages over the non-ACC ethylene procedure: it greatly improves the sensitivity of the test by enhancing ethylene production, it permits detection of small differences in vigor levels and it allows detection of ethylene prior to germination within a few hours of soaking.
Gokhan Hacisalihoglu and Anwar A. Khan
The effects of chemical or physical factors during pregermination imbibition phase, or on dry seeds, on embryo growth potential (EGP) was studied in lettuce (Grand Rapids and Mesa 659) and tomato (H-9889) seeds in relation to dormancy, invigoration, and vigor loss. Embryos were excised from treated seeds (washed if imbibed in chemical solutions) and their growth rate (a measure of EGP) followed at 25°C at high magnification (X55). Treated seeds were also germinated at 25°C. In lettuce seeds, dormancy inducing treatments, i.e., a 2-day dark soak at 25°C with 50–100 μM tetcyclacis (TCY) or a 2-day dark soak in water at 35°C, reduced the subsequent embryo growth and germination rate at 25°C. The reduction was prevented by 1 mM GA4+7 or irradiation applied during dormancy induction. A -d osmoconditioning (OC) at 15C with -1.2 MPa PEG-8000 solution in light or in dark with added GA4+7 enhanced the EGP; addition of TCY reduced the EGP and the TCY inhibition reversed by GA4+7. A progressive reduction in EGP occurred with increase in vigor loss. In tomato seeds, a soak with 100 μM TCY in light or dark for 2 days at 30°C induced a dormancy, but had little effect on EGP. Application of GA4+7 plus TCY prevented dormancy induction without affecting EGP. A 4-day matriconditioning (MC) at 25°C in light or dark with moist Micro-Cel E enhanced the EGP; TCY and/or GA added during MC, had little effect on EGP. EGP progressively decreased as the aging period increased. Thus, in lettuce, the EGP is coupled with the reversible –GA/+GA or phytochrome-controlled dormancy induction/release process, enabling germination, its inhibition, or its enhancement. In tomato, the EGP is not subject to light or GA control. Reduction in EGP, accompanying vigor loss in both seeds, is independent of light or GA action.
Claudinei Andreoli and Anwar A. Khan
The level of 1-aminocyclopropane-1-carboxylic acid (ACC) was 0.55 nmol.g-1 in dry lettuce (Lactuca sativa cv. Emperor) seeds. After 4h soak at 25°, 35° and 35°C+ KIN (kinetin, 50μM), the levels were 0, 0.2 and 1.14 nmol.g-1 seeds, respectively. The level of ACC was higher at 35°+KIN than at 35°C for up to 16h soak. No ACC was detectable at 25°C during 2 to 16h soak. In the presence of 50μM ABA, ACC level decreased to 0.2 nmol.g-1 at 4h soak and to zero level during 8 to 16h soak. The level of l-(malonylamino) cyclopropane-1-carboxylic acid (MACC), in dry seeds was 14 nmol.g-1. Exposure to 35°C in the presence or absence of KIN increased the level to 40-42 nmol.g-1 within 2h soaking, while at 25° only a slight increase (23 nmol.g-1) occurred. As in the case of ACC, the level of MACC was higher at 35°C+ KIN than at 35° or 25° for up to 16h soak.When seeds were soaked in ABA, the pattern of MACC produced was similar to that produced at 35°C. The results indicate that ACC synthase activity is enhanced by the addition of KIN at 35°C resulting in increased synthesis and/or accumulation of ACC and MACC. The relationship of ethylene biosynthesis to changes during stress imposition and alleviation by various factors will be discussed.
Anwar A. Khan and Claudinei Andreoli
Dormancy was induced in nondormant (germinate readily in light or darkness) seeds of several lettuce cultivars (Mesa 659, Emperor, Empress, Montello, Ithaca) by soaking in the dark in 5-100μM tetcyclacis (TCY) for 24h at 25°C as the seeds failed to germinate in the dark upon removal of TCY by washing. Higher concentrations of TCY was needed to induce dormancy in the light Paclobutrazol (PP 333) was relatively less effective. No dormancy was induced in nondormant lettuce seeds soaked for 24h in 100μM ABA as the seeds germinated readily in the dark upon removal of the inhibitor by washing. Thus, contrary to popular belief ABA does not appear to be a dormancy factor. Dormancy induced by TCY was released by soaking seeds in petri plates in water at 25°C in the light, in the presence of 0.001-1mM GA4+7, or by moist-chilling for 4-15d at 5°C. Dormancy was also released when dried dormant (dormancy induced by TCY) Mesa 659 or Emperor lettuce seeds were planted in a moist peat-lite mix in plastic containers and kept moist for 30d at 5°C, as indicated by emergence of normal, healthy seedlings upon transfer of the containers to 25°C. The significance of TCY induced dormancy in altering planting strategy in field plantings of lettuce and other crops will be discussed.