Optimal pollination environment of tetraploid ginger (Zingiber officinale Roscoe) evaluated by in vitro pollen germination and pollen tube growth in styles

Pollen germination percentages in vitro of a tetraploid ginger (Zingiber officinale Roscoe), ‘4×Sanshu’, tended to be highest at around 20°C. Pollen tube growth in the styles was greatly enhanced at 17°C, i.e., pollen tubes penetrated into the entire stylar length in 66.7% of the styles used. Pollen stored for at least 3 h under 40–80% relative humidity (RH) almost completely lost its germinability, whereas pollen incubated for up to 3 h under 100% RH retained a relatively higher germinability. Moreover, the pollen germinability that was lowered under 40–80% RH did not recover when the pollen was re-humidified for 2 h under 100% RH before inoculation onto a medium. When stigmas were pollinated with fresh pollen at 17°C under 100% RH, pollen tubes penetrated into the entire stylar length in 26.7% of the styles tested. Under 40–80% RH, however, pollen tubes stopped growing at the middle or the base of styles. From these results, I concluded that the optimal pollination environment of the tetraploid ginger was at 17–20°C under 100% RH.     

Molecular identification and antibiotic control of bacterial contamination in cultures of ginger (Zingiber officinale)

Zingiber officinale, Rosc., one of the major tropical spices in the world, belongs to the family Zingiberaceae. Bacterial contamination is a major constraint on the cryopreservation of in vitro shoot tip explants. The main objective of this study was to identify the nature of this bacterial contamination and its response to various antimicrobial agents (e.g. the antibiotics cephotaxime and streptomycin sulphate, or copper sulphate) for more effective control. The bacteria isolated from ginger plantlets were identified by amplification and sequencing of their 16S ribosomal DNA, followed by partial sequence analysis. Luteibacter yeojuensis was found in all contaminated cultures. L. yeojuensis is found mainly in soil and as a human pathogen. We believe this is the first report of L. yeojuensis contaminating in vitro cultures of ginger. Among the antimicrobial agents tested in the shoot multiplication medium [i.e. 1.0× Murashige and Skoog salts + 2.5 mg l^–l 6-benzylaminopurine + 3% (w/v) sucrose + 0.45% (w/v) Clarigar^TM], 100 mg l^–1 cephotaxime was most effective at reducing bacterial growth. It also gave the maximum number of shoots per shoot bud explant compared to the same medium supplemented with streptomycin sulphate, or CuSO₄, or control medium. No further bacterial contamination was observed when 8-week-old cultures were then subcultured on the same medium without added antibiotic or CuSO₄.     

姜四倍体离体诱导及其形态学分析

以丛生芽为材料,在离体培养条件下比较了秋水仙素不同浓度、不同处理时间诱导姜体细胞染色体加倍的效果。结果表明:以0.20%秋水仙素液体培养基处理8d的诱导效果最好,诱变率和成活率分别达到了42.86%和70.00%。根尖染色体压片检查发现,四倍体染色体数为2n=4x=44,二倍体对照为2n=2x=22。四倍体与二倍体相比表现出植株高大,茎秆变粗,叶片长、宽、厚均增大,叶片下表皮气孔密度减小,气孔和保卫细胞增大,保卫细胞内叶绿体数增加等特征,其中气孔密度及保卫细胞内的叶绿体数可作为鉴别四倍体与二倍体的重要性状。     

Antioxidant phytochemicals and gingerol content in diploid and tetraploid clones of ginger (Zingiber officinale Roscoe)

Fresh rhizomes of 18 diploid and tetraploid genotypes of Indian ginger, including commercial cultivars and experimental genotypes, were assayed for gingerol content and antioxidant activity. The tetraploid ginger type was derived from the respective diploid ginger by shoot tip culture. [6]-Gingerol was identified as the major pungent phenolic compound in all samples (mean 1354.78 ± 470 μg/g), while [8]- (mean 180.94 ± 45 μg/g) and [10]-gingerol (mean 234 ± 79 μg/g) occurred in lower concentrations. The total gingerol content of the tetraploid type was much higher than that of the respective diploid type and especially the [10]-gingerol. Both diploid and tetraploid types of the cv. Meghalaya Local contained the highest level of gingerol. At both ploidy levels, the genotypes showed significant differences in phenols and antioxidant activities. Correlation of total phenolics with different antioxidant assay at both diploid and tetraploid level was linear and strong to moderate. For genotypes at both ploidy levels, antioxidant activity (BCO) was determined especially by [6]-gingerol while [8]- and [10]-gingerol did not affect BCO. These results demonstrated that in the tetraploid types of ginger, the gingerol content and antioxidant activity were higher than in the diploid ones.     

山楂胚离体培养及四倍体诱导研究

以山楂为材料,研究了幼胚和成熟胚离体培养及四倍体诱导技术。花后40 d的山楂幼胚离体培养的成苗率很低,供试的15份资源的成苗率都小于20%。成熟胚离体培养的成苗率较高,调查的4份资源的成苗率均大于50%。用0.5%秋水仙碱+1%二甲基亚砜的混合溶液处理山楂成熟胚48 h,然后在SC+IBA 0.1 mg.L-1+TDZ 1.0 mg.L-1培养基上诱导芽分化。对秋水仙碱处理成熟胚再生植株进行茎尖染色体数目观察,检测的67个隆化粉肉植株中有3个是四倍体,变异率达4.5%。研究建立了山楂多倍体离体诱导体系,为培育山楂多倍体品种奠定了重要基础。     

Meristem culture and micropropagation of a variety of ginger (Zingiber officinale Rosc.) with a high yield of oleoresin

Summary

Meristems of ginger with or without leaf primordia were induced to form shoots on three-quarter strength Murashige-Skoog’s (MS) medium containing sucrose 6%, coconut milk (CM) 20%, ascorbic acid (AA) 100 mg l^?1, glutamine (GL) 400 mg l^?1, activated charcoal (AC) 250 mg l^?1, 6-benzylaminopurine (BAP) 0.5 mg l^?1, indolebutyric acid (IBA) 0.4 mg l^?1 and agar 0.8%. Meristem-derived shoots exhibited consistent multiplication on three-quarter strength MS medium containing sucrose (3%), AA (100 mg l^?1), AC (100 mg l^?1), BAP (4–5 mg l^?1) and agar (0.8%). Liquid media (agitated or static) were less effective than a solid (agar-gelled) medium for micropropagation. Kinetin and naphthalene acetic acid (NAA) incorporated at various levels (0.01–0.8 mg l^?1) with or without added BAP and IBA neither improved plantlet formation nor enhanced shoot multiplication. The in vitro plants were successfully established in vivo and the rhizome yield was comparable with that of plants grown by conventional methods.     

姜组织培养研究进展

对姜的器官(营养芽、花芽、花药)培养、细胞悬浮培养、原生质体培养和植株再生等方面的国内外研究现状进行了综述,并分析了姜组织培养研究中的主要问题。     

寒富苹果叶片离体再生及四倍体诱导

为建立寒富苹果高效的离体再生体系和多倍体诱导体系,以试管苗叶片为外植体,研究了培养基中激素对寒富叶片再生芽的影响及适宜的四倍体诱导方法。结果表明,当培养基中BA质量浓度为0.5mg/L时,再生频率最高达35.7%;当培养基中BA质量浓度为2.5mg/L时,再生频率超过90%,平均再生芽数在4以上。在培养基中附加1.0mg/LTDZ,再生频率达100%,平均再生芽数达19.47。以附加15、30、60、120mg/L秋水仙素的液体再生培养基处理叶片5d,各个质量浓度处理均诱导出四倍体植株,诱变率在5.3%～22.2%之间;寒富苹果叶片在附加50mg/L秋水仙素固体再生培养基上处理5d亦获得了四倍体植株。研究结果表明寒富苹果叶片具有极强的不定芽再生能力,叶片再生过程中进行秋水仙素处理是获得其四倍体的一个有效途径。     

姜的研究进展

对国内外姜的资源与分类、栽培现状、组织培养和应用价值等的研究进展进行了综述,详细讨论了外植体、激素、培养基等因素对姜组织培养的影响,以及组织培养过程中的体细胞无性系变异问题.并对今后姜的主要研究和应用方向进行了展望.     

Effect of photoperiod on flower bud initiation and development in myoga (Zingiber mioga Roscoe)

Selection of suitable production locations in Australia and New Zealand for production of myoga (Zingiber mioga Roscoe) has been limited by lack of information on climatic influences on flowering. This study focused on photoperiod as potential production sites within Australia differ considerably in daylength due to the geographical range. The two cultivars available in Australia (Inferior and Superior) were examined in this trial due to previously observed differences in vegetative and reproductive development.Plants grown under long-day conditions (16 h) and short-day conditions (8 h) with a night break produced flower buds, while those under short-day conditions (8 h) did not. The failure of plants under short-day conditions to produce flower buds was due to abortion of developing floral primordia rather than a failure to initiate inflorescences. It was concluded that for flower development in myoga a qualitative long-day requirement must be satisfied, but that flower initiation was day-neutral.Short-day conditions resulted in abortion of flower primordia, premature senescence of foliage and reduced foliage dry weight in both cultivars. Early senescence and low flower bud yield of the Inferior cultivar, but not the Superior cultivar have been observed in crop evaluation trials in Southern Australia and New Zealand. Differences in critical photoperiod between the two cultivars may explain this observation and therefore photoperiodic requirements may be an important consideration in site and planting date selection for different cultivars.     

Modeling individual leaf area of ginger (Zingiber officinale Roscoe) using leaf length and width

Leaf area estimation is an important biometrical observation one has to do for comparing plant growth in field and pot experiments. In this study, a leaf area estimation model was developed for ginger (Zingiber officinale Roscoe), using linear measurements of leaf length (L) and maximum width (W). Leaves from five ginger varieties (Varada, Rejatha, Mahima, Maran and Himachal) were used to develop the model in 2006–2007. The actual leaf area (LA) was measured with a leaf area meter (LI-3100, LI-COR, Lincoln, NE, USA) and taken as reference LA. The linear measurements were used to build linear (LA = a + b × L × W) and power models (LA = α × (L × W)^β) for each variety, as the modeling among variety were not different from each other, data for all five varieties have been pooled and compared with earlier models by graphical procedures and statistical criteria such as Mean Square Error (MSE), Root Mean Square Error (RMSE) and Chi-square (χ²). The selected model was validated during 2007–2008. The validation data set was used to produce a validation model for each variety by re-estimating the model parameters to develop the estimation model and the models were compared for consistency. The predicted LA (PLA) was compared with observed LA (OLA) by graphical procedures and lack of agreement was evaluated by calculating the relative bias, estimated by the mean of differences (d) and the standard deviation (SD) of the differences. Normality test was carried out by Spearman's rank correlation coefficient (r_s) and residuals were normally distributed. Finally, the proposed model for leaf area estimation of ginger is LA = −0.0146 + 0.6621 × L × W, R² = 0.997. This model can be reliably used for estimating leaf area of ginger non-destructively. The same equation can be extrapolated to all varieties and land races of ginger as it is vegetatively propagated crop with narrow genetic variability.     

姜种质资源离体保存技术研究

采用正交实验设计,研究了生长调节素、渗透压、温度等因素对姜种质资源离体保存的影响,以期建立姜种质资源离体保存技术体系,并通过生长恢复与遗传稳定性考察对保存材料进行初步评价.结果表明:生长调节剂、渗透压、温度等因素对姜种质资源的离体保存均有显著影响,最终确定姜离体保存的最佳保存方法为:MS+蔗糖60 g/L+琼脂4.0 g/L+甘露醇10 g/L+矮壮素(CCC)2.0 mg/L+多效唑(PP333)2.0 mg/L+脱落酸(ABA)2.0 mg/L,培养条件为光照时间14～16 h/d,光照强度1 500 lx,温度20℃,在此条件下保存300 d,存活率在50％以上,保存材料生长恢复情况良好,染色体保持稳定.     

四川‘竹根姜’胚性细胞悬浮系的建立与植株再生

 选用四川省地方品种‘竹根姜’的茎尖组织建立了体细胞胚性细胞悬浮系, 并通过悬浮培养获得了姜再生植株。结果表明悬浮细胞干质量受pH影响。接种量和AgNO₃ 对悬浮细胞的生长都有影响:最佳接种量为1.0% , 最适AgNO₃浓度为6.0 mg·L ^{- 1}。胚性细胞悬浮系增殖后, 接种到前期筛选的成熟分化培养基上获得了再生植株。     

中国砂梨多倍体种质资源发掘及其花粉育性分析

【目的】发掘中国砂梨种质资源中蕴藏的多倍体种质,明晰中国砂梨基于染色体水平的多样性,为砂梨多倍体育种和起源进化等研究提供依据。【方法】采用流式细胞术较为系统全面地对国家果树种质武昌砂梨圃中收集保存的466份砂梨种质进行倍性鉴定,并对新发掘的多倍体砂梨种质进行花粉生活力和自交结实率分析。【结果】共发掘出多倍体砂梨种质5份且均为三倍体,占总样本的1.07%,显示中国砂梨种内基于染色体的多样性水平较低。三倍体种质的花粉生活力和自交结实率均为0,显示其花粉不育且无单性结实能力。【结论】首次将流式细胞仪成功地用于中国砂梨种质资源的染色体倍性分析,发掘出的5份三倍体种质中,有3份自然三倍体集中分布于鄂西恩施的秦巴山区和武陵山区结合部,暗示该区域或为砂梨的起源演化中心之一。花粉育性分析结果可为其在多倍体育种等研究中的应用提供参考。     

In vitro polyploid induction of orchids using oryzalin

A protocol for in vitro polyploid induction using oryzalin was developed for Dendrobium, Epidendrum, Odontioda, and Phalaenopsis orchids (Orchidaceae). Protocorms and protocorm-like bodies (PLBs) in liquid nutrient media were subjected to oryzalin treatments of 14.4, 28.9, and 57.7 μM (w/v) concentrations for 3 and 6 days. Higher concentrations and longer treatment durations lowered the survival rates of the explants, but increased the number of polyploids produced. Stomatal guard cell lengths as measured with digital image analysis software, helped to identify several polypoids from digital images of leaf imprints. The optimal treatments were: 14.4 μM for 6 days in Dendrobium and Odontioda; 57.7 μM for 6 days in Epidendrum; and, 14.4 μM for 3 days in Phalaenopsis.     

Establishment and plant regeneration of somatic embryogenic cell suspension cultures of the Zingiber officinale Rosc.

Somatic embryogenic cell suspension cultures of four ginger cultivars were established. Somatic embryogenic calli were induced from ginger shoot tips on MS agar medium supplemented with 1.0 mg l⁻¹ 2,4-D and 0.2 mg l⁻¹ Kn, which contained only half concentration of NH₄NO₃. Rapid-growing and well-dispersed suspension cultures were established by subculturing this kind of callus in the same liquid MSN medium. The suspension cultures (about 1–2 mm in diameter) were placed on the MSN agar medium for callus proliferation. Thereafter embryogenic callus (1.5 cm²) was transferred to solid media (MS + 0.2 mg l⁻¹ 2,4-D + 5.0 mg l⁻¹ BA + 3% sucrose + 0.7% agar). Somatic embryos produced shoots and roots, and shoots developed into complete plantlets on solid MS medium supplemented with 3.0 mg l⁻¹ BA and 0.1 mg l⁻¹ NAA. The relationship between the DW of suspension cultures and pH changes in medium is also discussed. The suspension cultures still kept their vitalities after subculture for 8 months.     

生姜茎尖组织培养和污染率控制的研究

以生姜茎尖为外植体,筛选了愈伤组织和芽分化的最适培养基和培养方式,并探讨了生姜组织培养过程中污染率的控制方法.试验结果表明,将表面消毒与抗生素浸泡处理相结合,能有效降低污染率.培养基MS 6-BA 1 mg/L IAA 1 mg/L是愈伤组织诱导与芽分化的最适培养基.     

In vitro propagation of Blandfordia grandiflora (Liliaceae)

Multiple shoots were obtained when single shoot tips of Blandfordia grandiflora were exposed to different concentrations of kinetin, N6~ benzylaminopurine (BAP) and N6- isopentenylaminopurine (2iP). The best multiplication results were obtained when explants were subjected to kinetin at 8 (iM BAP at 0.5,2 and 8 (iM and 2iP at 2,8,32 and 128 |iM. Preliminary rooting trials were performed with three different auxins: indo- lebutyric acid (IB A), naphthalene acetic acid (NAA) and indole acetic acid (I A A). IB A at a concentration of 8 |iM and IAA at a concentration of 32 |xM gave highest root number, but when transplanted to the glasshouse, the best surviving plantlets were those treated with 2 |xM of IBA or 0.5 μM of IAA.     

生姜施用钢渣硅肥防病增产的效果试验初探

山东莱芜是"中国生姜之乡",生姜种植面积达1万hm2之多,年产量30多万t,但近年来种植户偏重施化肥,有机肥投入减少,加之生姜在该区连年种植,导致土壤营养比例严重失调,生姜病虫害加剧.     

In vitro micrografting and its applications to fruit science

Improvements have been brought to in vitro micrografting. The breaking of dormancy by dipping the seeds of the rootstock in concentrated solutions of benzylaminopurine avoids having recourse to long stratification in the cold (60–120 days according to species). The initial pre-treatment of apices with cytokinin allows their development and facilitates later grafting. Moreover, adding DIECA to the cut sections avoids oxidation and, together with the insertion of a piece of agar between apex and rootstock, increases the success of micrografting. Finally, a special technique of in vivo micrografting of pretreated apices on young plants has been developed.Micrografting is commonly used to obtain peach trees free of Sharka and NRS viruses, and to study incompatibility mechanisms in heterografts, leading to an early diagnosis of incompatibilities.