Somatic embryogenesis and plantlet regeneration from encapsulated embryos of Selinum tenuifolium

Summary

This paper reports, for the first time, somatic embryogenesis and synthetic seed production in Selinum tenuifolium Wall. Mature leaf explants inoculated in Murishige and Skoog (MS) medium supplemented with 3 µM 2,4-dichlorophenoxyacetic acid (2,4-D), containing 3% (w/v) sucrose and 0.7% (w/v) agar, induced 67% callus. Maximum production of globular structures, their differentiation into embryos and germination, occurred with a combination of 2 µM benzyladenine (BA) and 2 µM indole-3-butyric acid (IBA). To protect somatic embryos and produce synthetic seeds, gel capsules were standardised using a combination of sodium alginate and calcium nitrate concentrations. Gel capsules were most effective when formed with a combination of 3% (w/v) sodium alginate and 100 mM calcium nitrate for 30 min. The addition of MS medium to alginate capsules with 3% (w/v) sodium alginate, 3% (w/v) sucrose, 2 µM BA and 2 µM IBA significantly improved their germination rate to 77.8%, as well as their resulting shoot length (5.6 cm) and root length (7.2 cm), compared to controls (57.8%). Most plantlets (66%) survived under nursery condition. Storage at 4°C for different periods (10 d or 20 d) significantly (P < 0.05) reduced the percentage survival and germination of somatic embryos and artificial seeds compared to controls or 5 d storage.     

Influence of calcium propionate on the control of post-harvest rots of jujube and the biocontrol activity of an antagonistic yeast

The effect of calcium propionate (CaP) on post-harvest pathogens of winter jujube (Zizyphus jujuba Mill.), and on improving the efficacy of the biocontrol yeast, Metschnikowia pulcherrima, were evaluated. In vitro, CaP at ≥2% (w/v) had significant inhibitory effects on spore germination and germ-tube growth in Penicillium citrinum and Alternaria alternata. However, the application of 2% (w/v) CaP did not affect population growth of M. pulcherrima in nutrient yeast dextrose in vivo. With or without CaP, yeast cell numbers increased after 72 h by approx. 11.1-fold and 13.4-fold, respectively. The concentration of the antagonistic yeast suspension influenced the incidence of both pathogens and lesion development on jujube fruit, and the effect increased with an increase in yeast cell concentration. A combination of the antagonistic yeast plus 2% (w/v) CaP resulted in a lower incidence of disease and smaller lesion diameters compared to treatments with the antagonistic yeast alone. In this case, the incidence of disease was reduced by 16.8–27.3% for P. citrinum and by 25.2–31.4% for A. alternata. Lesion diameters on these fruit were 2.7–6.2 cm and 1.8–2.4 cm for P. citrinum and A. alternata, respectively. Calcium propionate had a consistent ability to enhance the biocontrol activity of M. pulcherrima against blue mould (P. citrinum) and Alternaria rot on jujube fruit. Thus, the application of 2% (w/v) CaP could be a useful approach to improve the efficacy of the antagonistic yeasts used for post-harvest disease control in jujube.     

In vitro plantlet regeneration from protocorms of Dendrobium lituiflorum Lindl. and Cymbidium bicolor Lindl. and their acclimatisation: effect of salts,sucrose, and banana extract

Summary

The salts present in modified Knudson C (KC) medium (1946), 2% (w/v) sucrose, and a natural banana extract (BE) [1 – 20% (v/v)] were studied singly, or in combination, for their effects on the regeneration of protocorms of the orchid species, Dendrobium lituiflorum and Cymbidium bicolor. The incorporation of KC salts along with higher percentages of BE [10% or 20% (v/v)] promoted protocorm development as well as shoot and root formation. The presence of 2% (w/v) sucrose in the medium further enhanced root lengths and the rooting percentage, irrespective of the presence of KC salts. Incorporation of both KC salts and 2% (w/v) sucrose was found to be obligatory, in conjunction with BE (a cost-effective natural additive) for plantlet regeneration. In vitro-raised plantlets of both orchid species exhibited high rates of survival under greenhouse conditions. Scanning electron microscopy revealed elliptical stomata and epicuticular granular wax deposits on their abaxial leaf surfaces after acclimatisation.     

对羟基苯甲酸对豇豆枯萎病菌的抑制作用

研究了不同浓度的对羟基苯甲酸(p-HB)对豇豆枯萎病菌孢子萌发、菌丝生长及挑战接种后对幼苗发病率的影响。结果表明：低浓度p-HB对豇豆枯萎病菌菌丝生长抑制作用不明显,而中、高浓度（5.0、10.0 mmol·L^-1）p-HB对菌丝生长的抑制作用达到极显著水平;在1.0～10.0mmol·L^-1范围内,p-HB 随着浓度的升高对豇豆枯萎病菌孢子萌发的抑制作用越大;1.0～10.0 mmol·L^-1的p-HB均能降低幼苗的发病率,尤其是浓度为10.0 mmol·L^-1时,能显著降低幼苗的发病率。     

Biological control of post-harvest anthracnose rot of loquat fruit by Pichia membranefaciens

Summary

The effect of Pichia membranefaciens on reducing post-harvest anthracnose rot caused by Colletotrichum acutatum in loquat fruit was investigated. Fruit were treated with different suspensions of P. membranefaciens before challenge with 1.0 10⁵ spores ml^–1 of C. acutatum, then incubated at 20°C for 6 d. The results showed that 1 10⁸ colony-forming units (CFU) ml^–1 of washed cell suspensions of the yeast provided better control of anthracnose rot than yeast in culture broth at the same concentration. Treatment with autoclaved cell cultures or culture filtrates did not prevent the incidence of disease. The concentration of a washed cell suspension of P. membranefaciens had a significant effect on the efficacy of controlling anthracnose rot in loquat fruit. At a concentration range from 1 10⁶ to 1 10⁹ CFU ml^–1, the higher the concentration of the antagonist, the lower the incidence of the disease and the smaller the diameter of the lesions. In inoculated wounds of loquat fruit, populations of P. membranefaciens increased approx. 34-fold after 6 d of incubation at 20°C.Washed cell suspensions of P. membranefaciens also significantly inhibited spore germination and germ-tube growth of C. acutatum, in vitro. These results suggest that P. membranefaciens has potential as a biocontrol agent to reduce post-harvest decay caused by C. acutatum in loquat fruit.     

多粘类芽孢杆菌HK18-8对辣椒炭疽病菌的抑制作用及其定殖能力

从健康的化香(Platycarya strobilacea Sieb. et Zucc.)根际土壤中分离获得生防菌HK18-8,为探究其对辣椒炭疽病的生防潜力,通过对峙培养和孢子萌发试验发现,HK18-8显著抑制短尖刺盘孢(Collectotrichum acutatum)的菌丝生长和分生孢子萌发,其抑菌带达到10mm,孢子萌发抑制率为70.15%;进行离体辣椒果实接种试验表明,HK18-8菌悬液对辣椒炭疽病的防治效果达到88.58%,其预防效果高于治疗效果,HK18-8的发酵液和菌悬液的防病效果显著;采用平板稀释法测定其在辣椒果实的定殖能力,处理9 d时仍能保持10~6 cfu·g~(-1)以上;根据形态特征、生理生化特性及16S rRNA序列同源性,HK18-8菌株鉴定为多粘类芽孢杆菌(Paenibacillus polymyxa)。     

Rapid regeneration of plants of Dendrobium lituiflorum Lindl. (Orchidaceae) by using banana extract

Effects of banana extract (BE) and 6-benzylaminopurine (BAP) were evaluated on asymbiotic seed germination and an early differentiation of protocorms and plant regeneration of Dendrobium lituiflorum Lindl. High percentage germination was achieved by culturing seeds on modified Knudson C medium supplemented with 10% (v/v) BE. Rapid regeneration was observed within 60 days of culture on 10% (v/v) BE supplemented KC medium where maximum percentage propagules showed development of leaves and root formation. Propagules on BAP supplemented KC medium showed no further development beyond one leaf stage. In another experiment, culture of shoots on 12.5% (v/v) BE supplemented KC medium led to multiplication, shoot elongation as well as vigorous rooting. Shoots cultured on 10 μM BAP supplemented MS medium showed maximum multiplication but these were stunted. Plants with well expanded deep green leaves and elongated roots from BE media were first hardened in vitro followed by ex vitro hardening on cocopeat:perlite (9:1) in the greenhouse conditions and exhibited 90% survival. The study emphasizes the role of BE as a natural additive at different stages of development from seed germination to plant regeneration.     

Effects of Artificial Pollination Using Pollen Suspension Spray on Nut and Kernel Quality of Pistachio Cultivar Owhadi

Yield and nut quality in pistachio are highly dependent on cultural practices especially pollination. This study was conducted to investigate the effects of pollen suspension spray containing 0.01% and 0.02% boric acid combined with 1% (w/v) pollen on nut and kernel quality of pistachio (Pistacia vera L. cv. Owhadi). The experiment was carried out as RCBD with three replications over 2011–12 period. The results showed that application of medium containing agar (A) and 0.01% boric acid (B1) combined with 1% (w/v) pollen decreased fruit set in both years compared to control whereas nut splitting was increased during 2012. Pollen suspension sprays unaffected nut and kernel dimensions in both years but increased kernel proteins and decreased oil during 2011. According to this study, it could be postulated that artificial pollination in pistachio using pollen suspension spray is possible.     

10种化合物对香蕉枯萎病菌的抑菌作用及对毒素钝化的效果

以香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)及其产生的毒素为供试材料,研究10种化合物在4种浓度下对香蕉枯萎病菌菌丝体生长、分生孢子萌发的抑制作用和对香蕉枯萎病菌毒素的钝化作用。结果表明,除NH4NO3外,供试化合物对香蕉枯萎病菌的菌丝体生长、分生孢子萌发及毒素均有不同程度的抑制作用,CuSO4在2～5g/L、FeSO4-7H2O在5g/L质量浓度范围内对菌丝的抑制效果最强,抑制率为100%。EDTA·Na2在5g/L质量浓度范围内对菌丝的抑制率为93.64%。FeSO4-7H2O、(NH4)2SO4-FeSO4-6H2O、KMnO4、CuSO4在0.5￣5g/L质量浓度范围内对分生孢子有明显的抑制效果,平均抑制率分别为97.73%、97.07%、90.98%、90.39%。CuSO4和KI在1￣5g/L的高质量浓度下均出现化合物中毒现象,在0.5g/L的质量浓度下钝化效果最好,分别达75%、67.05%。EDTA·Na2、Zn-SO4-7H2O和(NH4)2SO4-FeSO4-6H2O是在5g/L质量浓度下钝化效果最强,分别为100%,98%和93.99%。     

Effects of development,temperature, and calcium hypochlorite treatment on in vitro germinability of Phalaenopsis seeds

There are no standardized procedures for sanitizing orchid seeds for propagation by tissue culture and there is insufficient information about the optimum stage of orchid seed development for best germination. Phalaenopsis amabilis flowers were hand-pollinated and fruits harvested 90, 105, and 120 d after pollination (DAP) for seed developmental analysis. Embryo cell number per seed was counted after staining with 4′-6-diamidino-2-phenylindole and viewing through a confocal microscope. Germination percentage and cell number per embryo increased from 14 to 61% and 41 to 66%, respectively, during fruit development from 90 to 120 DAP. Seeds from mature, browning (∼140 DAP) Phalaenopsis Sogo Lit-Angel and Phalaenopsis spp. breeding line 9450 seed pods failed to germinate until frozen at −196, −80, or −18 °C and thawed or chilled at 4 °C for 10 d. Germinability in 140 DAP seeds was correlated with cracked testa after freezing and thawing. P. amabilis seeds were treated with 0, 5, 10, or 15% calcium hypochlorite (CH) for 5, 10, or 15 min. Ninety six percent of untreated seeds from 90 DAP fruit produced protocorms within 40 d after sowing (DAS). Exposing seeds to 5% CH for 10 or 15 min decreased germination to 85 and 73%, respectively. Exposure to 10 or 15% CH for 5, 10, or 15 min produced seed germination percentages of less than 40%. Protocorms developed root hairs and shoot primordia by 50 DAS and an average of one leaf and root by 85 DAS after treatment with either 0 or 5% CH. Higher concentrations delayed or inhibited protocorm development. Green fruits 120 DAP produced the highest percentage of protocorms, while ∼140 DAP seeds from browning fruit were dormant but cold treatments increased germination.     

Factors affecting the efficacy of agar-based substrates for the study of tomato pollen germination

Summary

The present work examines the effects of substrate additions and incubation conditions on in vitro germination of tomato pollen on semi-solid, agar-based substrates with the ultimate aim of defining a standard semi-solid substrate for tomato pollen germination studies. Partial replacement of sucrose by polyethylene glycol (PEG-6000) for osmotic regulation of the substrate significantly increased pollen germination of both the low temperature-susceptible F₁ cultivar ‘Dombito’, and the more temperature-resistant cv.‘Supermarmande’, to a level that varied according to the season of pollen harvest. In contrast, partial substitution of sucrose by mannitol was inhibitory to an extent that depended on the final concentration of mannitol in the medium. The optimum pH for germination was 6.5 and the optimum incubation temperature was 15°C. Among the vitamins (riboflavin, thiamine, pyridoxine, niacin, pantothenic acid, p-aminobenzoic acid), amino acids (glutamic acid, aspartic acid), casein hydrolysate, plant growth regulators (gibberellic acid, indole-3-acetic acid, indole-3-butyric acid, indole-3-propionic acid, 1-naphthaleneacetic acid, kinetin) and flavonoids (naringin, myricetin, naringenin, quercetin, rutin) tested, a significant increase in pollen germination was induced only by the flavonoids quercetin and myricetin and, to a lesser extent, by indole-3-acetic acid. In contrast, both kinetin and naringenin were inhibitory, as were both amino acids at 100 mg l^–1 and casein hydrolysate (at 1.0 – 10.0 g l^–1). On the basis of the above results, we conclude that, for both tomato cultivars tested, the most suitable semi-solid substrate among those examined was that containing 10% (w/v) sucrose, 15.1% (w/v) PEG-6000 and 1.5% (w/v) agar, with the possible addition of 5 mg l^–1 quercetin, or 5 mg l^–1 myricetin.     

The improvement of seedling production in two subfertile raspberry crosses following the application of growth substances

The effect of exogenous growth substances on the yield of seedlings, on some aspects of fruit development, on seed and embryo growth and on germination was studied in two crosses, 4n M. Jewel selfed and 4n M. Jewel χ 2n M. Jewel, following treatment of the young fruits with combinations of gibberellic acid (GA), para-chlorophenoxyacetic acid (CPA) and 2-naphthoxyacetic acid (NOA) 2, 4, 6, 8 and 10 days after pollination. The treatments affected the set of large pyrenes, seed size and percentage germination but had no effect on embryo size or on the percentage of pyrenes containing seeds. The number of seedlings obtained was affected both by set and by percentage germination. Seed size was positively correlated with percentage germination in the 4nx2n cross only. A mixture of CPA and NOA increased seedling yield in both crosses.     

Changes in antioxidants and fruit quality in hot water-treated ‘Hom Thong’ banana fruit during storage

The effects of hot water treatment on antioxidants and fruit quality were investigated in banana fruit of cv. Gros Michel (Musa acuminata, AAA Group, locally called cv. Hom Thong) by immersing fruits in hot water (50 °C) for 10 min, before storage at 25 °C for 10 days or 14 °C for the first 8 days followed by storage at 25 °C for the second 8 days until ripening. Quality parameters including peel color and pulp firmness indicated that hot water treatment helped to delay banana fruit ripening at both storage conditions. Hot water treatment decreased the levels of hydrogen peroxide (H₂O₂) and malonydialdehyde (MDA) during storage at 25 °C. Glutathione (GSH and GSSG) contents and the ratio of GSH/GSSG during fruit approaching ripening were significantly induced in hot water-treated fruits while ascorbic acid (AA) contents were slightly increased. In addition, the combined treatment increased free phenolics and flavonoids during storage. Results suggest that hot water treatment has led to an induction of antioxidants in banana fruits as indicated by an increase of antioxidants and a decrease of H₂O₂ during ripening, and all of which result in a delayed ripening of banana fruit.     

柑橘炭疽病的病原鉴定和综合防治措施

采集桔园感病柑橘,进行病原菌分离鉴定并研究了其对农药的敏感性、形态学和ITS1-5.8S rDNA-ITS2序列分析。结果表明:病原菌为胶孢炭疽菌（Colletotrichum gloeosporioides）;对11种市售农药都具有敏感性,其中氟硅唑、中生菌素、多抗霉素、丙森锌、氟菌唑、异菌脲6种药物对病菌菌丝生长的抑制活性较高,以氟硅唑最为灵敏,其EC₅₀为0.041μg/mL,丙森锌和中生菌素次之,EC₅₀分别为1.59、4.73μg/mL;氟硅唑和百菌清对孢子的抑制效果较好,丙森锌、佳爽和多抗霉素也有一定的保护作用。根据药敏试验和文献方法制定了综合防治方案。     

Relations of Phenolic Inhibitors to Resistance of Immature Apple Fruits to Rot

Extracts of phenolic compounds from maturing Red Delicious apple tissues containing p-coumaryl-quinic acids and chlorogenic acid were shown to be inhibitory to germination of Botrytis cinerea spores and mycelial growth of B. cinerea, Penicillium expansum and Alternaria sp. Measurements of the content of both acids during the growing season showed that the content of phenolic compounds was highest early in the season and decreased as the fruit matured. The decrease coincided with the rise in susceptibility of the fruit to rot caused by B. cinerea. The concentration of p-coumaryl-quinic acid was highest near the skin and lowest near the core.     

Selection and testing of epiphytic yeasts to control anthacnose in post-harvest of papaya fruit

Anthracnose, caused by Colletotrichum gloeosporioides, is a major post-harvest disease in papaya fruit. The major objectives of the present work were to isolate, select and test the in vitro and in vivo ability of epiphytic microorganisms, isolated from papaya fruit and leaf surfaces, in controlling anthracnose onset after harvest. A total of 75 bacteria, 67 yeasts and 22 mycelial fungi were isolated. Thirty yeast isolates were able to inhibit the mycelial growth of C. gloeosporioide in vitro and seven of those were used in in vivo assays, resulting in the identification of two very effective isolates. Isolate CEN63, identified molecularly as Cryptococcus magnus, was the most effective in controlling the disease and therefore was studied in more detail. The results of the assays with C. magnus provided evidence that when fruit were treated with the antagonists at concentrations of 10⁷ to 10⁸ cells/ml, as early as 24 h, preferentially 48 h, before inoculation with the pathogen, the development of disease was significantly reduced. C. magnus is a potential antagonist for the development of a commercial product. Additional studies on the modes of action of this yeast isolate, as on its ability to interact with fungicides are being conducted to generate solid basis for the development of an environmentally friendly control agent.     

Biological control of the shoot disease of Amaranthus hybridus caused by Choanephora cucurbitarum with Bacillus subtilis

Summary

Isolates of Bacillus subtilis from soil and ogili (a local food condiment) controlled choanephora shoot disease of the vegetable crop plant, Amaranthus hybridus, in the greenhouse. Disease developed on plants inoculated simultaneously with the pathogen Choanephora cucurbitarum and the ‘ogili’ isolate, but not the soil isolate, of B. subtilis. Application of either bacterial strain to plants one day before a challenge with the pathogen prevented disease development. A single application of either strain prevented disease development from a subsequent challenge with the pathogen at any time over a thirty day period. Viable counts of the microflora on the shoot tips of the treated plants indicated strongly that the inoculated bacteria multiplied and colonized the extending shoot tip whether or not the shoots were covered with polythene bags. Both strains of Bacillus subtilis inhibited mycelial extension growth as well as conidial germination in vitro.     

GA和CPPU对枇杷无核果品质的影响

以６～１０ 年生的几个枇杷品种为试材,进行ＧＡ＋ ＣＰＰＵ 不同的浓度、使用时期和时期组合试验,结果表明：用ＧＡ１ ０００ ｍ ｇ·ｌ－ １处理花蕾,再用ＧＡ１ ０００ ｍ ｇ·ｌ－ １＋ ＣＰＰＵ２０ ｍ ｇ·ｌ－ １在花后１ 月下旬、２ 月下旬、３ 月下旬分期喷果３ 次,是增大无核果和防止无核果落果的重要措施;无核果率１００ ％ ,无核果单果重与对照正常果单果重差异不显著,其果肉厚、可食率均极显著高于对照,生理病果较少;可溶性固形物含量极显著低于对照,果形指数显著大于对照,成熟较迟。并探讨了上述处理所产生的无核果存在问题的对策     

Rapid in vitro germination of zygotic embryos of fluted pumpkin (Telfairia occidentalis Hook. f.)

Summary

Fluted pumpkin, Telfairia occidentalis, is becoming an important regional vegetable for its food and medicinal uses. The recalcitrant nature of its seed makes conservation difficult and in vitro techniques may be a viable option for conservation. A pilot study was conducted on the effects of different concentrations of a commercial bleach [3.85% (w/v) sodium hypochlorite] for surface sterilisation of T. occidentalis seed. The optimum concentration [25% (v/v)] was then used as a basis to investigate the responses of mature embryonic axes of T. occidentalis to different concentrations of kinetin (Kin; 0, 1.0, or 2.0 mg l^–1) and 1-naphthaleneacetic acid (NAA; 0, 0.5, or 1.0 mg l^–1) combined in a factorial design. The results of the first experiments indicated that commercial bleach at 25% (v/v) resulted in the lowest contamination of explants (10%), with no evident injury to the embryonic axes. The results revealed that root emergence started 3 d after initiation (DAI) only on Murashige and Skoog medium (MS) with no added plant growth regulator (PGR), and that, by 12 DAI, all media supported the rooting of explants. The highest rooting percentage (69%) was observed at 15 DAI on MS medium with 0.5 mg l^–1 NAA, without Kin. However, shoot emergence started at 9 DAI on PGR-free MS medium, on MS with 0.5 mg l^–1 NAA, or on MS plus 1.0 mg l^–1 Kin. The highest shooting percentage (91%) of explants was observed with 0.5 mg l^–1 NAA at 21 DAI. Considering all other growth parameters, MS medium supplemented with 0.5 mg l^–1 NAA was found to be best for the germination of embryonic axes of T. occidentalis.     

Studies on allelochemical and mineral compounds for sustainable weed control in a pavement filler from brick recycling material

Here, organic and inorganic compounds, most of them naturally occurring, were selected on the basis of their potentially high resistance to leaching, in order to explore their potential to prevent the germination of weed seeds in inert substrates as well as in pavement filler based on brick wall recycling material (BRC). Using the weed species Lolium perenne, Poa pratensis, and Plantago lanceolata, it was shown that the susceptibility to either alkaline or allelochemical environments was plant species-dependent with regard to seed germination. In a cumulative leaching test, where a total of 53 L of water was rinsed through 1 L of filler material, the organic compounds juglone, lactic acid, pelargonic acid, and pelargonaldehyde all lost their initial inhibitory effect within a short time. In contrast, the suppressive effect of 2% w/v boric acid and 20% w/v sodium silicate (with a molar ratio of Si/Na of 3.35) was as persistent as that achieved by the sodium silicate-enriched commercial joint filler Dansand®. Results reveal sodium silicate as the most promising tested compound to achieve a long-term weed-suppressive effect in BRC with the lowest potential harm to soil invertebrates.