Gibberellic acid and flower bud development in loquat (Eriobotrya japonica Lindl.)

The application of gibberellic acid (GA₃) to the whole loquat tree from mid-May to early June and from early August to the onset of flowering, significantly reduced the number of premature flowering shoots per current shoot and per m³ of canopy, and so reduced the total number of panicles per m³ of canopy. The number of vegetative shoots per m³ of canopy was also significantly reduced by applying GA₃. The response depended on the concentration applied and produced optimal results at 50 mg l⁻¹. Differences in the number of flowers per panicle and leaves per shoot were not significantly modified by the treatment. Nevertheless, GA₃ applied directly to the developing apex near to flower differentiation reduced the number of flowers per panicle by 25–35% and without modifying the morphological characteristics of the panicle. Results suggest that less sprouting of lateral buds was largely responsible for the reduction in flowering intensity caused by GA₃. Best treatments reduced thinning costs (60%, approximately) of premature flowering shoots, slightly increased fruit diameter and significantly improved fruit colour and juice quality, thus advancing harvest date.     

In vitro germination of walnut (Juglans regia L.) embryos

The present studies were undertaken with a view to standardize the medium and culture conditions for embryo culture of five cultivars of walnut viz., ACO 38853, Netar Akhrot, Gobind, Solding Selection and Blackmore. Embryos from mature fruits were aseptically excised and cultured on MS medium supplemented with different combinations of BAP, kinetin and GA₃. Best performing medium was MS with 0.5 mg l⁻¹ kinetin, 0.5 mg l⁻¹ BAP and 2 mg l⁻¹ GA₃ yielding 66.6% germination in Netar Akhrot after 12 days of culturing. Percent germination of excised embryos was higher when GA₃ and cold treatments were simultaneously applied as compared to those when applied separately. Netar Akhrot was found to be the best responding cultivar, which had a range of 25–66.6% embryo germination under different culture conditions. Plantlets with shoots and roots have been obtained in Netar Akhrot and ACO38853 and are transferred to soil after hardening.     

Improving ‘Bing’ sweet cherry fruit quality with plant growth regulators

Final fruit diameter is the prime determinant of sweet cherry fruit value. Previous research has shown that mesocarp cell size accounts predominantly for variability in final fruit size, within a genotype. Our research program evaluated the potential to improve sweet cherry fruit size/weight with growth regulators to affect cell division and/or cell expansion stages. In the current study we screened 8 plant growth regulators (PGRs), including cytokinins, gibberellins, and auxins, and their combinations for their ability to increase ‘Bing’ fruit weight. Each PGR was mixed in lanolin paste and applied to fruit pedicels at 9 or 30 days after full bloom (DAFB), to coincide with estimated peak in cell division and cell expansion activity, respectively. Several cytokinins applied 30 DAFB improved fruit weight significantly (ca. +15%) with N-(2-Chloro-4-pyridyl)-N′-phenylurea (CPPU) and 6-(3-hydroxybenzylamino) purine (mt-Topolin) at 100 mg l⁻¹ being the most effective. Gibberellins, applied alone, improved fruit size and delayed fruit maturation and exocarp coloration. GA₃ at 200 mg l⁻¹ applied at 9 DAFB was the most effective and improved final fruit weight by 15%. Fifty-six percent of the fruit from this treatment were ≥9 g compared to 15% of similar weight fruit from untreated limbs. Both GA₃ and GA_4/7 treatments applied 9 DAFB increased fruit radial expansion. 4-Chlorophenoxyacetic acid, a synthetic auxin, also stimulated higher fruit growth rates at stage I and stage II, and fruit color development, but did not improve final fruit size.     

Growth and flowering of black iris (Iris nigricans Dinsm.) following treatment with plant growth regulators

The effects of application method and concentration of gibberellic acid (GA₃), paclobutrazol and chlormequat on black iris performance were assessed. Plants (10 cm high, 4 ± 1 leaves) were sprayed with 125, 250, 375 or 500 mg L⁻¹ or drenched with 0.25, 0.5, 1 or 2 mg L⁻¹ GA₃. In a second experiment, the plants were sprayed with 100, 250, 500 or 1000 mg L⁻¹ or drenched with 0.25, 0.5, 1 or 2 mg L⁻¹ paclobutrazol. Other plants were sprayed with 250, 500, 1000 or 1500 mg L⁻¹ or drenched with 100, 250, 375 or 550 mg L⁻¹ chlormequat. In each experiment, the control treatment consisted of untreated plants. Results indicated that the tallest plants (37.3 cm) in the GA₃ experiment were those sprayed with 250 mg L⁻¹. The most rapid flowering (160 days after planting) occurred when a 375 mg L⁻¹ GA₃ spray was used, whereas flowering was delayed to 200 days using 1 mg L⁻¹ GA₃ drench. Drenching with 1 mg L⁻¹ GA₃ increased height of the flower stalk by 7 cm compared to the control. Though relatively slow to flower, plants drenched with 1 mg L⁻¹ GA₃ had long and rigid stalks, which were suitable as cut flowers. Number and characteristics of the sprouts were not affected by GA₃. All paclobutrazol sprays resulted in leaf falcation. A 500 or 1000 mg L⁻¹ paclobutrazol spray resulted in severe and undesirable control of plant height, drastic reduction in stalk height and weight, and delayed flowering. Plants drenched with 0.25 or 1 mg L⁻¹ paclobutrazol were suitable as pot plants. Chlormequat reduced plant height only at the highest drench concentration, which also reduced flowering to 70%. No leaf falcation was observed with GA₃ or chlormequat. Chemical names: ( ± )-(R*,R*)-beta-((4-chlorophenyl)methyl)-alpha-(1,1,-dimethylethyl)-1H-1,2,4,-triazol-1-ethanol (paclobutrazol); (2-chloroethyl) trimethylammonium chloride (chlormequat).     

Response of evergreen perennial tree crops to gibberellic acid is crop load-dependent. I: GA3 increases the yield of commercially valuable ‘Nules’ Clementine Mandarin fruit only in the off-crop year of an alternate bearing orchard

Worldwide, gibberellic acid (GA₃) is used routinely to increase fruit number and size of seedless mandarins. The efficacy of seven combinations of GA₃ concentrations and application times to maximize total yield and yield of commercially valuable fruit (diameter 57.2–76.2 mm) of ‘Nules’ Clementine mandarin (Citrus reticulata Blanco) was determined in a commercial orchard. GA₃ applied during the period of intense flower abscission failed to reduce the total number of abscised flowers in both the light off- and heavy on-bloom years. No GA₃ treatment reduced fruit abscission when trees were setting the low yield off-crop. However, all trees receiving GA₃ in the high yield on-crop year had fewer abscised fruit than untreated control trees (P = 0.0188) and GA₃ applied 10 days after 75% petal fall and in July increased the number of fruit retained on tagged branches >20% compared to control trees (P = 0.0005). Maximum air temperature was not related to flower or fruit abscission. In the off-crop year (548 fruit per untreated control tree), it was necessary to apply 15 or 25 mg L⁻¹ GA₃ at 60% bloom, 90% bloom, 75% petal fall and 10 days after 75% petal fall to significantly increase the number of fruit per tree and yield of commercially valuable fruit (kilograms and number per tree) (P < 0.0001) above that of control trees, with no reduction in total kilograms per tree. In the following on-crop year, it was better not to apply GA₃: no treatment increased total yield or fruit size and five of seven GA₃ treatments tested reduced total yield as kilograms and number of fruit per tree (P = 0.0003). The results provide strong evidence that GA₃ efficacy is crop load-dependent and dictate that crop load should be considered when using GA₃ to increase fruit set or fruit size of mandarins.     

CPPU application on size and quality of hardy kiwifruit

For the purpose of determining the appropriate conditions of application to increase the size of a hardy kiwifruit, Actinidia arguta ‘Mitsuko’, N₁-(2-chloro-4-pyridyl)-N₃-phenylurea (CPPU) was applied at three different growth stages of the crop: at petal fall, 10 and 25 days after petal fall (DAPF), and three different concentrations: 1, 5 and 10 mg l⁻¹. A significant increase in fruit size was obtained by treatment at the concentrations of 5–10 mg l⁻¹ and at 10 DAPF. The fruit weight doubled. Although a significant reduction in the concentrations of total soluble solids (TSS), titratable acids (TA) and ascorbic acid (AsA) in the CPPU-treated fruits was recorded, the TSS/TA ratio and AsA content per fruit increased by the treatment. CPPU application at petal fall induced abnormally protruding fruit tip.     

Influence of growth regulators on setting,retention and weight of fruits in two cultivars of litchi

Investigations were undertaken to explore the possibility of improving setting, retention and weight of fruits in ‘Early Seedless’ and ‘Calcuttia’ cultivars of lichi (Litchi chinensis) by means of growth regulators. Indole acetic acid (IAA) at 20, 40 and 80 mg l^?1, 2,4-dichlorophenoxy acetic acid (2,4-D) at 2,4 and 8 mg l^?1 and gibberellic acid (GA₃) at 50, 100 and 150 mg l^?1 were sprayed on panicles in the first fortnight of April, when 50–100% flowers had opened. All 3 growth regulators caused a favourable effect on fruit setting, fruit retention and weight of individual fruits, but IAA at 20 mg l^?1 proved the best for enhancing setting, GA₃ at 50 mg l^?1 for increasing retention and GA₃ at 100 mg l^?1 for improving fruit weight. IAA and GA₃ should, therefore, be used in combination. Between the 2 cultivars tested, ‘Calcuttia’ proved superior to ‘Early Seedless’ in fruit setting, fruit retention and weight of individual fruits.     

The inhibition of flowering by means of gibberellic acid application reduces the cost of hand thinning in Japanese plums (Prunus salicina Lindl.)

The application of gibberellic acid during flower bud induction significantly reduced flowering of ‘Black Diamond’ and ‘Black Gold’ Japanese plums. The response depended on the concentration applied and on the type of shoot. Mixed shoots had a similar response in both varieties, flowering being reduced by 40% for GA₃ 50 mg l⁻¹ and by 75–90% for GA₃ 75 mg l⁻¹ or higher concentration. With regard to spurs, GA₃ 50 mg l⁻¹ reduced flowering intensity by 40% and 25% in ‘Black Gold’ and ‘Black Diamond’, respectively, and GA₃ 75 mg l⁻¹ or higher concentration reduced flowering by 70% and 50%, respectively. This partial inhibition of flowering significantly reduced the cost of manual thinning. The best GA₃ concentration was found to be 50 mg l⁻¹, since it reduced the cost of thinning by 45–47% and increased final fruit weight by 7–33% for ‘Black Diamond’ and ‘Black Gold’, respectively. Not significant differences in yield and in mature fruit characteristics of treated trees were found compared to untreated trees.     

Production of inter-section hybrids between Primula filchnerae and P. sinensis through ovule culture

Inter-section hybrids were obtained in the reciprocal crosses between Primula filchnerae (2n = 2x = 24) of Sect. Pinnatae and P. sinensis ‘Fanfare’ (2n = 2x = 24) of Sect. Auganthus by rescuing ovules on half-strength (1/2) Murashige and Skoog's (MS) medium supplemented with 50 g l⁻¹ sucrose, 2.5 g l⁻¹ gellan gum, 0.1 mg l⁻¹ α-naphthaleneacetic acid (NAA), 0.1 mg l⁻¹ 6-benzyladenine (BA) and 50 mg l⁻¹ gibberellic acid (GA₃). In ovule culture, germination occurred with radicle elongation but no plumule was observed. The radicle kept on the initial medium showed root proliferation with callus formation. When the calluses were transferred to (1/2)MS media containing 30 g l⁻¹ sucrose and 3 g l⁻¹ gellan gum, without plant growth regulators (PGRs) or with 1 mg l⁻¹ zeatin and 0.1 mg l⁻¹ NAA, plantlets were regenerated. The plants thus obtained were confirmed to be hybrids through flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses. The hybrid obtained when P. filchnerae was used as the maternal parent was diploid, whereas hexaploid hybrid was obtained when using P. sinensis as the maternal parent. The hexaploid hybrid might be produced through chromosome doubling of a triploid originated from the fertilization of P. sinensis with unreduced pollen of P. filchnerae.     

Effect of synthetic auxins on fruit development of ‘Bing’ cherry (Prunus avium L.)

The main cherry cultivar grown in the warm climate of Israel, ‘Bing’, produces relatively small fruit. Over three consecutive years (2003–2005), application of 50 mg l⁻¹ 2,4-dichlorophenoxypropionic acid [2,4-DP; as its butoxyethyl ester (Power™)], 10 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid [3,5,6-TPA; as the free acid (Maxim^®)], or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) plus 30 mg l⁻¹ naphthaleneacetic acid (NAA; 0.3% Amigo™), at the beginning of pit-hardening when fruitlet diameter was ca. 13 mm caused appreciable and significant increases in fruit size and total yield, except when the crop load was heavy. Anatomical studies revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect on fruit quality, either at harvest or after 1 month of storage at 0 °C, or on return yield in the following year.     

Potential of ethephon,NAA, NAD and urea for thinning pistachio fruitlets

Alternate bearing, the occurrence of high yield ‘on’ year followed by low yield ‘off’ year, is striking in pistachio (Pistacia vera L.). Floral buds of pistachio are formed a year before bloom, but abscise during the years with heavy crop (‘on’ year). Abscission of floral buds is due to competition between growing seeds on 1-year-old shoots and developing buds on current season growth. We studied the effects of chemical fruit thinning on alternate bearing and nut characteristics in a commercial orchard of ‘Owhadi’ pistachio cultivar during 2003–2004 in Rafsanjan, Iran. In both years, ethephon (100 and 200 mg l⁻¹), naphthaleneacetic acid (NAA) (125 and 250 mg l⁻¹) and urea (2.5 and 5%) were applied to the branch units of each individual ‘on’-year trees. The results showed that ethephon at both concentrations significantly increased fruit thinning and floral bud retention for the subsequent year. Other treatments also increased fruit thinning and floral bud retention but were inferior to ethephon.     

Clonal propagation of Rosa clinophylla Thory. through axillary bud culture

A protocol for clonal propagation of Rosa clinophylla, a rare and endangered species but very important for breeding purposes had been standardized through in vitro axillary bud culture. Although cytokinins alone were able to induce shoot buds, but their proper growth and number could be increased only when they were used in combination with GA₃. However, there was shoot tip necrosis and leaf fall in the proliferated shoots. AgNO₃ at 58.85 μM proved effective to avoid shoot necrosis and yellowing of leaves. Activated charcoal (AC) at 250 mg l⁻¹ was found necessary at all the stages of shoot multiplication as well as rooting. Ninety percent rooting could be achieved in 1/2 MS medium supplemented with 4.92 μM IBA and 250 mg l⁻¹ AC. Rooted plantlets were hardened and transferred to the field successfully with 80% survival rate.     

Effect of synthetic auxins on fruit size of five cultivars of Japanese plum (Prunus salicina Lindl.)

Most of the Japanese plum (Prunus salicina) cultivars grown in Israel produce relatively small fruit. Application of 2 l solution tree⁻¹ of 25 mg l⁻¹ 2,4-dichlorophenoxypropionic acid (2,4-DP) as butoxyethyl ester (Power™), 15 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid (3,5,6-TPA) as free acid (Maxim^®), or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) + 30 mg l⁻¹ naphthaleneacetic acid (NAA) (0.3% Amigo™) at the beginning of pit-hardening, when fruitlet diameter was ca. 22 mm, caused an appreciable and significant increase in fruit size. The yield of large fruit per cv.: ‘Kesselmen’ (100% increase), ‘Songold’ (100%), ‘Black Diamond’ (800%), ‘Royal Diamond’ (160%) and ‘Royal Zee’ (100%). As a result, the total yield of all five cultivars was also increased dramatically. Anatomical studies with ‘Songold’ revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect either on fruit quality at harvest (and after 1 week in shelf-life), or on return yield in the following year.     

Effects of exogenous application of plant growth regulators on the development of ovule and subsequent embryo rescue of stenospermic grape (Vitis vinifera L.)

Seedless grapevine cultivars (Vitis vinifera L.) are widely grown in Europe, America and Asia. Abortion of zygotic embryos in seedless grapes largely limits the efficiency of breeding of seedless cultivars through genetic crossing. The present study was designed to investigate effects of exogenously applied plant growth regulators (PGRs) to the grapevines in field condition on ovule and subsequent embryo rescue of seedless grapes of small seed traces. First experiment was performed by measuring ovules weight, proportion of each category ovules in maturity and embryo development in vitro of seedless grape cv. Centennial Seedless, Thompson Seedless and Crimson Seedless sprayed by chlormequat (CCC), benzyladenine (BA), ethephon (CEPA) and putrescine (Put). The effects of different application concentration and date of CCC were further evaluated in Centennial Seedless in later experiment. The results showed that exogenous application of all PGRs did not affect the total number of ovules per berries in maturity. CCC increased the ovules weight and proportion of ovules >4 mm in length of three varieties in maturity. The effects of two application times of PGRs on weight of berries and ovules and proportion of each category ovules in maturity were not significantly different. In the proceeding of embryo rescue, CCC at 100 and 1000 mg l⁻¹, BA at 100 mg l⁻¹ and Put at 20 mg l⁻¹ increased the percentage of developed embryos of Centennial Seedless and Thompson Seedless. The results showed that the size of ovules excised for embryo rescue significantly affected embryo formation and plant regeneration. The percentage of embryos formation in ovules >4 mm in length was significantly more than in ovules 2–4 mm in length, no embryo was found in ovules <2 mm in length. Exogenous application of CCC at 100–500 mg l⁻¹ significantly increased percentage of ovules >2 mm in length by 80.0–82.7% in Centennial Seedless, therefore improving embryo formation. The statistical correlation was found between the proportion of ovules >2 mm and embryo formation (r = 0.92) in Centennial Seedless. Among the different spraying time in Centennial Seedless, CCC applied 14 days before bloom produced significantly more ovules >2 mm in length and embryos formation.     

Plant regeneration of pansy (Viola wittrockiana) ‘Caidie’ via petiole-derived callus

An in vitro plant regeneration protocol for pansy (Viola wittrockiana) cultivar ‘Caidie’ from petioles was established as following: callus induction on a half-strength MS medium supplemented with 0.45 μmol l⁻¹ 2,4-d plus 8.9 μmol l⁻¹ BA, callus subculture on medium F (1/2MS with 4.5 μmol l⁻¹ 2,4-d, 2.7 μmol l⁻¹ NAA and 0.44 μmol l⁻¹ BA) and then on medium T (1/2MS with 4.5 μmol l⁻¹ 2,4-d, 2.7 μmol l⁻¹ NAA and 2.2 μmol l⁻¹ BA), shoot regeneration on medium D3 (MS media supplemented with 2.9 μmol l⁻¹GA₃, 23.6 μmol l⁻¹ AgNO₃, 0.02% active charcoal and 4.5 μmol l⁻¹ TDZ), shoot multiplication on medium M (half-strength MS medium containing NAA 1.1 μmol l⁻¹, TDZ 9.1 μmol l⁻¹ and GA₃ 8.7 μmol l⁻¹), and then shoot elongation and rooting on medium R (MS medium supplemented with 1.1 μmol l⁻¹ NAA and 1.1 μmol l⁻¹ BA). Subculture on appropriate medium was found to be important for successful shoot regeneration.     

Recurrent somatic embryogenesis and plant regeneration in Coriandrum sativum L.

An efficient method of repetitive somatic embryogenesis and plant regeneration was established in Coriandrum sativum L. Embryogenic callus was induced from cotyledon and hypocotyl segments on Murashige and Skoog (MS) medium with 4.52 μM 2,4-dichlorophenoxy acetic acid (2,4-D), upon subculturing on medium having same level of 2,4-D at an interval of 3 weeks developed somatic embryos, which progressed to cotyledonary stage through early developmental stages of somatic embryogenesis. The transfer of somatic embryos at an early cotyledonary and cotyledonary stage in clumps in succession to fresh 4.52 μM 2,4-D supplemented medium developed embryos in a cyclic manner. Upon transferal to embryogenic clumps (cotyledonary embryos) to modified MS medium (4 g l⁻¹ KNO₃, 0.29 g l⁻¹ NH₄NO₃, 3 mg l⁻¹ thiamine HCl, 0.5 mg l⁻¹ pyridoxine HCl, and 5 mg l⁻¹ nicotinic acid), the embryos irrespective of the cycles underwent maturation and germination. Germinating embryos transferred to half-strength MS medium favored healthy growth of plantlets. The system of recurrent somatic embryogenesis in coriander offers a system for genes transfer and also scale-up production of modified plants.     

Micropropagation of the bromeliad Guzmania ‘Hilda’ via organogenesis and the effect of α-naphthaleneacetic acid on plantlet elongation

This study established a highly effective micropropagation system to obtain good plantlet proliferation from floral organs via callus induction and bud differentiation in Guzmania ‘Hilda’ bromeliad. The best frequencies of organogenic callus formation (20% in petal and 35% in ovary explants) were obtained on media containing a combination of 1.0 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) with 1.0 mg l⁻¹ α-naphthaleneacetic acid (NAA) and 1.5 mg l⁻¹ 2,4-D with 0.5 mg l⁻¹ NAA, respectively. Organogenic calli were cultured on medium with 1.0 mg l⁻¹ NAA and 0.5 mg l⁻¹ 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (TDZ) induce the differentiation and regeneration of adventitious buds into plantlets. When the plantlets were cultured in a medium with optimum NAA concentration (0.5–1.0 mg l⁻¹) significant improvement in regeneration and elongation was achieved within one month. This overcame the difficulty of delayed elongation in Guzmania plantlets. More than 99% of the regenerated and acclimatized plantlets developed to the flowering stage.     

Molecular characterization and analysis of somaclonal variation in chrysanthemum cultivars using RAPD markers

Molecular characterization using RAPD analysis was carried out in eight cut flowers and two pot plant cultivars of chrysanthemum. Three of them (‘Refocus’, ‘Red Reagan’, and ‘Sheena Select’) were established in vitro and the occurrence of somaclonal variation was studied using the same molecular technique. Two induction media (MS + 0.1 mg l⁻¹ NAA + 0.1 mg l⁻¹ BA, and MS + 2.0 mg l⁻¹ IAA + 0.5 mg l⁻¹ Kinetin), and two proliferation media (MS + 0.1 mg l⁻¹ NAA + 0.2 mg l⁻¹ BA, and MS + 4.0 mg l⁻¹ IAA + 2.0 mg l⁻¹ Kinetin) were employed in order to evaluate the effect of the medium composition in the shoots’ stability. Likewise, the effect of the culture age was considered in assessing genetic stability. Monthly subcultures were carried out, identifying the origin and history of the shoots, throughout a nine-month proliferation period followed by acclimatization. Molecular markers were obtained in every subculture cycle and from the acclimatized plants. Only one shoot from the 7th subculture of the cultivar ‘Refocus’ showed a different band pattern. The use of RAPD for chrysanthemum cultivar characterization and somaclonal variation detection is discussed.     

Brassinosteroid analogue effects on the yield of yellow passion fruit plants (Passiflora edulis f. flavicarpa)

The effects of successive brassinosteroid analogue (BR) applications (0.1 mg l⁻¹ of brassinosteroid analogue BB-16) were evaluated on commercial yellow passion fruit (Passiflora edulis f. flavicarpa) orchards in the first year of production. The treatments applied were: control, BR-1 (1 BR application shortly after the first flowers appeared), BR-2, BR-3, BR-4 and BR-5 (BR application in two, three, four and five consecutive weeks after the appearance of the first flowers, respectively). The fruits were collected for seven consecutive weeks (105 fruits treatment⁻¹) and fruit mass, length and diameter, soluble solid contents; pulp mass and peel thickness were evaluated in the laboratory. Multivariate analysis was performed in order to determine whether there were differences among the treatments taking into account all the measurements made. BR-3 was the most promising treatment because it produced the highest number of fruits plant⁻¹ (81.5) compared to the control (53.5) and the soluble solid content was 1 °Brix greater than the control. The BR-3 treatment resulted in a 65% increase in the estimated yield of the passion fruit plants, corresponding to 20 t ha⁻¹ compared to the control yield of 12 t ha⁻¹. The results showed that BB-16 sprayed during a period of reproductive development can increase the number of fruits per plant.     

In vitro rescue of immature avocado (Persea americana Mill.) embryos

An in vitro culture protocol was developed as a means of avocado embryo rescue. Different factors including presence of cotyledons, medium texture and cold or gibberellic acid pretreatments, were studied. To better understand the germination process in this recalcitrant species, immature zygotic embryos at different stages were used in these experiments. Optimum results were dependant on the embryo developmental stage. Whereas smaller embryos (5 mm long) germinated better in M1 liquid medium, 15 mm long embryos responded better when precultured in B5m medium supplemented with 1 mg l⁻¹ GA₃, and fully mature embryos were capable of germinating directly in solid M1 medium. Our results suggest the existence of two types of dormancy in avocado embryos: an embryo-dormancy caused by cotyledons, and another type of dormancy, mainly occurring in 35 mm long embryos and revealed by the formation of dwarfing rosette seedlings, that can be released by a GA₃ pretreatment.