Effects of Four Culture Media on Callus Induction Rate of Wheat Anther

Effects of four culture media including MS, N6, C17 and K on wheat anther callus induction in vitro culture were studied. The results showed that the callus in- duction rate of four kinds of culture medium was in the order of K〉C17〉N6〉MS.

Validating field regeneration capacity for selected accessions of Gossypium hirsutum using callus induction and regeneration capacity

Background Gossypium hirsutum undergoes rapid clonal propagation to regenerate a mature plant through tissue culture.However,the correlation between cotton leaf regeneration,callus induction,and regeneration ability was still obscure.In this research,cotton leaf regeneration level for 21 accessions in the field(new leaves)was observed after the first harvest,and a comparison between field regeneration level and callus induction with its regeneration capacity(new shoots and roots)for the same 21 accessions was carried out.Agronomic traits,including plant height,leaf area,fresh leaf weight,dry leaf weight,the number of flowers and bolls,and biochemical(proline content)and physiological(chlorophyll and carotenoid content)traits during the flowering stage of 21 upland cotton accessions,were investigated.Result A significant correlation between physiological parameters and callus induction was discovered.Callus induction and regeneration capacity of roots and shoots for hypocotyl,cotyledons,and shoot tip tissues were used to validate field leaf regeneration level after the first harvest.CCRI 24 showed significant leaf regeneration in the field and callus induction capacity through callus induction and regeneration.Conclusion We found a substantial relationship between field regeneration capability and callus induction with its regeneration capacity for the hypocotyl,cotyledons,and shoot tip.The results showed that ZS061,Lumian 378,Jimian 863,and ZS065 have the highest moisture retention capacity,while CCRI 24,Liaoyang Duomaomian,and Beizhe Gongshemian have the lowest moisture retention capacity.CCRI 24 has the highest leaf regeneration capacity in the field,while Beizhe Gongshemian has the lowest leaf regeneration capacity.All our result provides a clue for checking the regeneration capacity through leaf regeneration level in the field.

Influences of Carbon Sources and Plant Growth Regulators on Anther Culture Efficiency of Pepper

[Objective]In order to increase anther culture efficiency of pepper.[Method]MS culture media and Bolajiaohong were used in this experiment to study the influences of carbon sources and concentrations on anther callus induction of pepper.Jiayu was taken as a material to study influences of plant growth regulators and concentrations on anther callus induction of pepper according to L16（4^5） orthogonal design.[Result]The average callus and embryoid induction rates of maltose at all concentrations were higher than these of sucrose but the difference was not significant.Taking maltose or sucrose as a carbon source,3% to 6% concentration was good for increasing induction frequencies of calli and embryoids.However,If the concentration was over 6%,the induction rates were declined dramatically with the increase of sugar concentration.The influences of growth regulators on induction rate of calli were listed as 2,4-D﹥ZT﹥NAA﹥KT﹥6-BA;the influences on induction rates of embryoids were listed as 2,4-D﹥NAA﹥ZT﹥KT﹥6-BA.The 2,4-D,ZT,NAA and KT had signficant or extremely significant influences on induction rates of calli and embryoids.2,4-D,ZT at 1.0 mg/L and NNA,KT at 0.5 mg/L had the best effects.The influences of ZT on calli and embryoids were better than those of KT and 6-BA.1.0 mg/L 2,4-D ＋1.0 mg/L ZT ＋0.5 mg/L KT ＋0.5 mg/L 6-BA was the best regulator combination for induction culture of Jiayu anther.[Conclusion]The experiment provided research basis for anther culture of pepper.

Preliminary Study on the Anther Culture in Lycium barbarum L.

To investigate the culture technique in anther of Chinese wolfberry,we optimized the culture medium(including hormone combination)and culture conditions.The results showed that calluses were induced from all the six tested Chinese wolfberry materials,but the induction rate of callus varied toward the materials with different genotypes.When the experimental materials were cultured on medium appended with 2,4-D 1.0 mg/L and KT 1.0 mg/L under dark,the callus induction rate reached 20.0 % in this study,and this hormone combination should be the optimum for anther culture of Chinese wolfberry.With MS appended with 6-BA 0.5 mg/L and NAA 0.1 mg/L as differentiation medium and that appended with NAA 0.1 mg/L,the plants could be yielded in 20 days.

Differences in Primary Callus Induction among Different Anthurium andraeanum Varieties

[Objective] This research aimed to optimize continuously the highly efficient regeneration system of Anthurium andraeanum. [Method] The leaves and petioles of four A. andraeanum varieties were used as explants to investigate the differences in primary callus induction among different A. andraeanum varieties. [Result]The callus formation capacity of SAM and SST was stronger than that of SDM and SHG. Among the four varieties, the leaf regeneration capacity of SAM, SDM and SHG was stronger than the corresponding petiole regeneration capacity. However,the petiole regeneration capacity of SST was stronger. The optimum medium for petiole callus induction of SST was 1/2 MS ＋ TDZ 4.0 mg/L ＋ 2, 4-D 0.2 mg/L with induction rate of 87.5%; the optimum medium for leaf callus induction of SAM was 1/2 MS ＋ TDZ 2.0 mg/L ＋ 2, 4-D 0.2 mg/L with induction rate more than90%; the optimum medium for leaf callus induction of SDM and SHG was all 1/2MS ＋ ZT 2.0 mg/L ＋ 2, 4-D 0.2 mg/L with induction rates of 59.34% and 79.63%,respectively. [Conclusion] In addition to variety differences, the differences in differentiation ability among different types of calluses should be also taken into account in the establishment and optimization of tissue culture and rapid propagation technology system of A. andraeanum.

Optimization of the Callus Induction System of Chenopodium quinoa Willd

Callus induction effects of nine varieties of Chenopodium quinoa Willd. were compared by taking stem segments and cotyledons of C. quinoa as the ex- plants. At the same time, callus JnductJon of stem segments was optimized, as well as the callus proliferation system. Research results showed that the optimal explant for callus induction was stem segment. The average callus induction rate of nine varieties reached 90% in culture medium MS ＋ 0.5 mg/L 2, 4-D. In the callus opti- mization test, treatment VI （MS ＋ 0.5 mg/L 2, 4-D ＋ 0.5 mg/L KT ＋ 0.5 mg/L NAA） and treatment II （MS ＋ 0.5 mg/L 2, 4-D） had close induction rate, but the callus morphology was greatly different. The latter had loose, glossy and yellowish white calluses. Therefore, culture medium MS ＋ 0.5 mg/L 2, 4-D was the optimal for callus induction. And using 2, 4-D together with KT and NAA could significantly increase the proliferation rate of calluses.

Factors Affecting Embryogenic Callus Production and Plant Regeneration in Anther Culture of Bupleurum chinense

Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense.Methods The different effects such as four genotypes,plant growth regulators,and temperature condition were compared in the experiments.The histological study was performed with the process of the anther culture.Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao(ZCYH),Z4,and Z5 at the early-to middle-uninucleate stages,except for genotype ZPM1 at the tetrad stage.Cold pretreatment increased the production of the embryogenic callus,in which 4-day cold pretreatment improved the production of embryogenic callus from 0% to 2.2% and 5.0% for genotypes ZPM1 and ZCYH,respectively.No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D) .The highest regeneration rate(34.6%) was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine(BA) .The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis.Chromosome counting of regenerated plantlets showed mostly diploid plant(2n = 12) with only one haploid plant(n = 6) .Because of the low rate of microspore embryo formation,we only tracked the process of embryogenesis from the connective tissue,instead of microspore by histological observations.Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system.This is the first report on the haploid plantlet through the anther culture of B.chinense.

Preliminary Study on Anther in vitro Culture of Lily“Prato”

The effects of the microspore developmental stage,hormones and culture condition on anther in vitro culture of lily（Lilium spp.） were discussed.The results showed that when the flower buds were about 23-26 mm long,the microspores were at the uninucleate stage which was suitable for culture and the culture under the darkness would promote the callus induction of anther.The induction frequency could reach 42.5% in the optimized medium which was MS＋[6-BA（0.5）＋KT（2.0）＋2,4-D（1.0）] mg·L-1.The rate of callus differentiation could reach 31.57% in the optimized medium which was MS＋ NAA（1.5,2.0） mg·L-1.

Effect of Different Treatment on Anther Callus Formation and Browning in Balsam Pear （Mornordica charantia L.）

Brown callus derived from anther limited the application of anther culture in balsam pear. In order to establish a more perfect regeneration system from anther cultuer, the effects of low temperature pretreatment, 2,4-dichlorophenoxyacetic acid （2,4-D）, vitamin C （Vc） and silver nitrate （AgNO3） on callus induction and browning in anther culture of balsam pear （Momordiea charantia L.） were investigated. The results showed that after pretreatment at 4 ℃ for 1 day, callus induction rate was the highest and browning rate was the lowest. Anthers on MS medium supplemented with 2,4-D 0.5 mg/L formed more and better callus. The medium supplemented with Vc or AgNO3 was advantageous to the induction of callus and reduction of browning. When cultured on medium supplemented with 50 mg/L Vc or 5 mg/L AgNO3, callus induction rate was the highest and browning rate was rather low.

Establishment and Optimization Growth of Shoot Buds-Derived Callus and Suspension Cell Cultures of <i>Kaempferia parviflora</i>

Callus and suspension cells culture of Kaempferia parviflora was successfully established. Meristematic shoots can be used for utilization of plant cell biosynthetic capabilities for obtaining useful products from valuable medicinal plant to meet out the pharmaceutical demand and also for studying the metabolism. The medium containing combination of 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.2 mg/L napthyleneacetic acid (NAA) promoted the highest callus induction at 20%. Transferring the initiated callus on the medium with 1 mg/L 2,4-D enhanced the proliferation rate up to maximum fresh weight of 6.71 gm. Growth curve of cultured cells revealed that the cells continued to grow until 50 days of culture and showed the highest peak (fresh weight) at 40 days in all different initial weight tested ( 0.2, 0.5 and 1.0 gram). Isolated embryogenic callus was found to produce the highest in weight when suspended in liquid medium supplemented with 1 mg/L 2,4-D at 110 rpm resulted 13.5 gram fresh weight and 1080 mg dry weight.

Effects of Genotypes and Basic Medium on Culture of Maize Mature Embryos

[Objective]This study was to screen out suitable genotypes and basic medium for the culture of maize mature embryos.[Method]Using mature embryos of nine maize genotypes as explants,the effects of genotypes and basic medium on callus induction and subculture were investigated.[Result]The genotypes performed better in callus induction and subculture were found in turn 853-35,853-209,Dan 34 and 81162.MS medium is better than N6 medium in the callus induction from maize embryos,while N6 medium is more suitable for callus subculture.[Conclusion]Our study further improved the tissue culture system in maize with mature embryos as explants.

Tissue Culture System for Different Hybrid of Indica Rice

The effect of varieties and media compositions on callus induction from rice anther and subsequent plant regeneration was studied. The results showed that the callus induction was not significantly different in different media, but mostly depended on genotypes; mean frequency of callus induction on F1 hybrid varieties showed that the medium supplemented with 100 mg·L^-1 concentration of the proline was useful for callus induction; when the anther derived callus sub-cultured on MS medium supplemented with BAP （0.5 mg·L^-1）, KT （1.0 mg·L^-1）, IAA （0.25mg·L^-1） and NAA （0.5 mg·L^-1） for plant regeneration, the frequency of regeneration ranged from 0 to 54.17%, and the highest differentiation index reached 30.5 after sub-cultured for three times; the plants were rooted after transferred to 1/2MS medium with NAA （0.5 mg·L^-1） and IAA （0.5mg·L^-1）.

Ag+及激素对藜麦愈伤诱导及分化的影响

本研究以‘津藜2号’藜麦(Chenopodium quinoa)种子为材料,采用MS培养基研究不同浓度Ag+、NAA和蔗糖对藜麦愈伤组织诱导和分化的影响。结果表明,在培养基MS+30 g/L蔗糖+20μmol/L Ag++0.8 mg/L NAA中藜麦愈伤组织诱导率最高,可达到100.0%。在培养基MS+40 g/L蔗糖+20μmol/L Ag++0.4 mg/L NAA中其愈伤组织诱导率最低,只有13.3%。在培养基MS+30 g/L蔗糖+20μmol/L Ag++1.5 mg/L 6-BA中其愈伤组织分化率最高,可达到100.0%。在培养基MS+20g/L蔗糖+20μmol/L Ag++2.0 mg/L 6-BA中其愈伤组织分化率最低,只有20.0%。藜麦最适愈伤组织诱导和分化培养基配方的筛选为其再生体系和转化体系的建立奠定基础,也为其种质资源保存、培育新品种以及工厂化育种提供科学依据。

百岁兰愈伤的诱导与悬浮细胞培养

为探究稳定高效的百岁兰叶片愈伤组织诱导体系及其悬浮细胞培养方法,利用百岁兰叶片作为材料,采用正交法使用两种激素(NAA与6-BA)配置植物培养基,运用避光与昼夜节律两种处理诱导愈伤组织产生,并利用愈伤组织与其对应配比的液体培养基培养悬浮细胞。结果表明,百岁兰叶片在0.5 mg/L 6-BA+1.0 mg/L NAA浓度下的诱导愈伤具有最好的稳定性与成功率,在昼夜节律条件下诱导与生长的效率很高,以此为基础培养的悬浮细胞取得了一定进展。研究可为百岁兰的保护与稳定遗传提供技术参考。

Advances in D. <i>melanoxylon</i>Investigations towards Tissue Culture: Problems and Limitations

The first attempt on D. melanoxylon tissue culture was conducted from 2010 to 2013 at a high level of expectations. A total of 500 seeds were sterilized at different concentration of reagents and inoculated at different strength of the Murashige and Skoog medium for germination to obtain disease free explants for callus induction trials. A total of 400 nodal segments obtained from germinated seeds were sterilized at different concentration of reagents and inoculated at different hormonal combinations to induce callus formation for seedling multiplication. Results from this tissue culture attempt set a foundation for tissue culture success in D. melanoxylon on the future research. Only 19.8% of seeds inoculated in half strength of Murashige and Skoog medium germinated within 7 days while only 6.8% of seeds inoculated in full strength of Murashige and Skoog medium germinated within 6 days. This germination was at sterilization of 20 minutes in 35% ethanol and 20 minutes in 2.6% sodium hypochlorite. A total of 1% of inoculated D. melanoxylon seedling fragments in Murashige and Skoog media supplemented with hormone combination at 2.0 mg/l BAP + 0.5 mg/l NAA developed callus after16 days from the inoculation day. The final weight of the callus at the last record was 0.62 g. In this induction ex-plants were surface sterilized in 35% ethanol for 20 minutes and 2.6% sodium hypochlorite solution for 20 minutes. The color of callus was green and friable in nature. Other hormonal combinations in this case did not induce callus production. These results suggested that the problems which affect seed germination in the natural environment are also reflected on germination in the Murashige and Skoog medium and in callus induction. Vulnerability to fungal attack is a limitation for successful callus induction and germination in the culture room. More research under improved sterile conditions is needed to improve callus percentage for seedling multiplication.

林木单倍体研究进展

结合国内外文献,论述了林木单倍体的研究进展,对林木单倍体诱导的几种途径进行对比总结,分析了目前在林木单倍体育种中存在的问题并对这些问题提出了建议,并对其广阔的应用前景进行了展望,以期为林木单倍体诱导研究提供科学依据。

不同IAA和6-BA质量浓度组合对甘薯茎尖培养和表型变异的影响

以甘薯品种烟薯29号的茎尖为外植体,MS培养基为基本培养基,研究不同质量浓度IAA(0.1、0.5 mg/L)和6-BA(0.1、0.5、1.0 mg/L)组合下愈伤组织诱导、芽分化、植株再生和表型变异情况。结果表明,MS培养基中IAA质量浓度为0.5 mg/L时对烟薯29号愈伤组织的诱导效果最佳,且利于茎尖成活,在此质量浓度下,平均愈伤组织诱导率和茎尖成活率分别达到了92.00%和88.00%。芽诱导率和丛生芽率随着培养基中6-BA质量浓度的升高总体上增加,在IAA质量浓度为0.1 mg/L的处理中,平均芽诱导率和丛生芽率分别达到了64.27%和80.56%。MS培养基中IAA质量浓度为0.5 mg/L时有利于植株再生,平均植株再生率达到了39.33%,在此质量浓度下,植株再生率和成苗数均随着6-BA质量浓度的升高而增加,成苗周期随着6-BA质量浓度的升高而缩短。烟薯29号的再生植株中出现了1株无性系变异植株,命名为YM6-11,其叶脉色、花冠形状和杂交亲和性与烟薯29号相比发生了改变。综上,烟薯29号茎尖培养最佳培养基组成为MS+0.5 mg/L IAA+1.0 mg/L 6-BA,将获得的再生植株用于实际生产时,应进行表型和遗传学鉴定,及早发现变异植株,减少不必要的经济损失。

Effects of CeCl_3 and LaCl_3 on callus and root induction and the physical response of tobacco tissue culture

La3＋ and Ce3＋ have positive effects on plant growth and production. Although it is well known that rare earth elements promote cell growth. The biological effects of La^（3＋） and Ce^（3＋） on callus, shoot and root induction in tobacco are still unclear. The relationships among callus induction, rooting, enzyme activities and stomatal characteristics in tobacco are unknown. The objectives of this study were to identify the relationships between the induction of calluses, shoots, roots, stomata and enzyme activities. The induction percentages of calluses, buds, roots were recorded at 5,10,15, 20 and 25 days after La^（3＋） and Ce^（3＋） treatments. Peroxidase isoenzyme activity was determined by electrophoresis. The characteristics of the stomata were observed under an optical microscope. Our results show that low concentrations of Ce^（3＋）（〈15 mg/L） result in increases in the induction percentages of calluses,buds and roots, but La^（3＋）（〉5 mg/L） inhibits the induction of calluses, buds and roots. There are more peroxidase isoenzyme bands in Ce^（3＋） treatments than in La^（3＋） treatments. This is consistent with the induction percentages of calluses,buds and roots in Ce^（3＋） and La^（3＋） treatments. High enzyme activities may promote the induction of calluses, buds and roots. The stomata area and stomata number of leaves are significantly different between La^（3＋） treatments and Ce^（3＋） treatments. La^（3＋） improves the stomata area and number. Based on these results, we speculate that La^（3＋） may promote the development of the photosynthetic system. Ce^（3＋）may promote tobacco growth and rooting by improving enzyme activities.

木蓝愈伤组织诱导与不定芽分化培养

初步建立了木蓝(Indigofera bungeana Walp.)的离体再生培养体系,为其基因工程育种研究提供前期技术参考。通过种子无菌播种技术获得野生木蓝无菌实生苗,以胚轴、茎段和叶片为外植体,筛选木蓝愈伤组织诱导、不定芽分化、增殖、壮苗培养的最适培养基配方。结果显示,木蓝种子无菌播种发芽率达98%,最适培养基为1/2MS。3种外植体在不同培养基下均能诱导出愈伤组织,根据愈伤组织生长及质地色泽筛选出最适诱导培养基,其中,叶片为MS+0.5 mg/L 6-BA+0.5 mg/L 2,4-D+0.5 mg/L NAA;茎段为MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+0.1 mg/L NAA;胚轴为MS+0.1 mg/L 6-BA+0.1 mg/L2,4-D+0.5 mg/L NAA。相比较叶片和茎段,胚轴来源的愈伤组织适于不定芽分化,最适分化培养基为MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+0.1 mg/L NAA和MS+2.0 mg/L 6-BA。不定芽增殖培养基为MS+1.0 mg/L 6-BA+0.1 mg/L NAA,壮苗和生根的最适培养基均为1/2MS。

不同LED光质配比和光照强度对红掌新品种福星组织培养的影响

【目的】光照条件是影响红掌组培效率的关键因素之一,研究不同LED光质配比和光照强度对红掌福星组织培养不同阶段的影响,探讨适宜红掌组培各阶段的LED光质配比和光照强度,为红掌高效组培提供重要技术参考。【方法】以红掌新品种福星为试验材料,设置不同比例的红光和蓝光LED光质配比和光照强度,分析其对红掌福星愈伤组织诱导、增殖培养及壮苗培养的影响。【结果】LED红光光质条件较适宜红掌福星愈伤组织诱导,光照强度为100 lx的10%R光质条件下,愈伤组织诱导率最高、达79.58%,且愈伤组织块呈黄绿色、紧实、体积大,状态表现好,适宜红掌福星叶片愈伤组织诱导;R∶B=3∶1光质配比适于红掌福星愈伤组织增殖培养,光照强度为600 lx时,增殖系数最高、达6.44;与WL光照条件相比,R∶B=4∶1光质配比更适宜红掌福星愈伤组织芽分化培养,愈伤组织芽分化在光照强度为1000 lx时,愈伤组织块的增殖分化芽个数较少、为19.44个,芽矮壮,无黄叶现象,生长势好;红掌福星壮苗阶段组培苗培养,适合的光质配比是R∶B=4∶1,光照强度为900 lx时,红掌福星壮苗植株根条数较多、平均为5.67条,株高2.88 cm、茎粗1.44 mm、叶长11.69mm、叶宽8.54 mm,植株叶片大,SPAD值为39.23,叶片叶绿素含量更高,植株整体生长健壮且无黄叶现象。【结论】LED光照条件对红掌福星愈伤组织诱导、增殖培养及壮苗培养均有影响,根据红掌组织培养不同阶段的光照需求,选用合适的LED光质配比和光照强度条件,能有效提高红掌组培愈伤组织诱导率、增殖率和组培苗生产质量,为红掌新品种福星高效标准化生产提供技术支持。