温度及培养基中添加物对辣椒花药培养胚状体诱导的影响

对11个辣椒基因型材料进行了花药培养研究,分别研究了温度处理、培养基中添加无机盐和激素对辣椒胚状体诱导的影响。结果表明:4℃低温预处理花蕾3~5 d和接种后33℃高温处理7~9 d均可大大提高胚状体诱导率;MS培养基中添加50μmol/L的AgNO3大幅度提高了胚状体诱导率。添加适量的激素也可以提高辣椒培养效果,其中,添加2,4-D或KT的最适浓度为0.1 mg/L,而添加较高浓度的NAA(1.0 mg/L)辣椒胚状体诱导率达到最高。     

Effects of colchicine and cold duration pretreatments on androgenesis responses of tomato (<Emphasis Type="Italic">Lycopersicon esculentum</Emphasis> Mill) via anther culture

The aim of the present study was to optimize the in vitro chromosome-doubling procedure by colchicine pretreatment in tomato anther culture. Anthers of 1.7–2.0 mm long were isolated at prophase-metaphase I and pretreated by different concentrations of colchicine (0, 250, 500 and 750 mg/l) at 4°C for 36 and 72 hrs under dark condition. After colchicine pretreatment, anthers were transferred to a colchicine free medium to callus and shoot induction. Analysis of variance showed that percentage of callus and shoot induction (PCI and PSI) were not influenced by cold pretreatment durations. However, significant differences were observed between cold durations for the number of regenerated (NRP) and the percentage of doubled haploid plants (PDH) (p < 0.05). Androgenesis responses were affected by different colchicine concentrations and interactive colchicine x cold durations (p < 0.05). Maximum PCI, PSI, NRP and PDH were belonged to 250 mg/l colchicine and 72 h cold duration. However, colchicine concentrations above than 250 mg/l had negative effect on androgenesis responses of anther cultures in both cold durations.     

甜(辣)椒花药培养胚状体诱导与植株再生

以10份不同基因型甜(辣)椒为试材进行花药培养,通过对基因型、取蕾时期、低温预处理、热激处理、碳源及外源激素浓度配比等因素的研究,建立有效的甜(辣)椒花药培养胚状体发生体系.结果表明:基因型是限制甜(辣)椒花药培养胚状体诱导的关键因素,不同品种间出胚率差异显著,其中品种003出胚率最高,为10.8%;处于盛花期的花蕾最适于甜(辣)椒花药培养;4℃低温预处理1-3 d有利于胚状体的诱导,以处理2 d的胚状体产率最高;以2%的麦芽糖代替3%的蔗糖能显著提高出胚率和子叶形胚的比率,筛选出适于甜(辣)椒花药培养胚状体诱导的最佳培养基为Ms+0.5 mg/L NAA+1.0 mg/L KT+2%麦芽糖,能有效地提高出胚率并促进植株冉牛;获得了6个基因型的子叶形胚和再生植株.     

影响辣椒花药胚状体发生的因素

对17个辣椒基因型材料进行了花药培养,分别研究了基因型、激素组合、高温预处理、接种时间对辣椒花药胚状体发生的影响。结果表明,不同基因型辣椒花药培养差异较大,其中,最易产生胚状体的是海花三号;添加合适的激素配比也可以提高辣椒花药培养效果,0.1 mg/L BA+0.5 mg/L NAA+1.0 mg/L KT是辣椒花药胚状体诱导培养最适宜的生长调节剂组合;接种后35℃高温预处理6~10 d可大大提高胚状体诱导率;2—3月接种的花药最易出现胚状体。     

Infection of Phytophthora capsici on pepper—Models and affecting factors

Under controlled conditions in a growth chamber, we studied the mortality of pepper seedlings caused by Phytophthora capsici. The results showed that soil temperature and soil water content were important factors affecting their infection with P. capsici and the optimum condition for infection was found to be a soil temperature of 22°C–28°C and a soil water content of 40%. The relationships of pepper seedling mortality caused by P. capsici along with soil temperature, soil water content and their interactions can be described by mathematical models. Field observations suggested that the Gompertz model was the best one for describing the epidemiological dynamics of the disease. The incidence of pepper phytophthora blight was significantly related to the initial incidence of pepper phytophthora blight, soil temperature, soil water content and air temperature. A forecasting model for pepper phytophthora blight in the field was developed. __________ Translated from Chinese Journal of Applied Ecology, 2007, 18(5): 1061–1065 [译自: 应用生态学报]     

Effect of dry and wet storage at cool temperatures on the post-harvest performance of Ranunculus asiaticus L. flowers

A study was undertaken to assess the effect of different storage temperatures on senescence and postharvest performance of isolated flowers of Ranunculus asiaticus L.. The main aim of the work was to develop a cost-effective storage protocol to reduce the postharvest losses and to bring out the transportation of cut flowers of R. Asiaticus. The flowers were subjected to two different storage treatments, dry storage and wet storage, and their postharvest performance was compared under laboratory conditions. For this purpose, the buds were harvested at 8:00 AM at loose bud stage (Stage II of flower development). The harvested buds were cut to a uniform size of 15 cm and processed for dry or wet storage. For dry storage, the buds were packed after wrapping them in moistened filter papers and kept at 5°C and 10°C. For wet storage, the buds were held in distilled water in separate glass beakers kept at 5°C and 10°C, respectively. A separate set of buds each for dry and wet storage was kept at room temperature (15±2)°C. After 72 h storage, the buds were kept at room temperature in distilled water. The average life of an individual flower that opened fully was about 4–5 days. The buds kept under wet storage at 5°C and 10°C for 72 h maintained their premature status, while the buds held at room temperature for 72 h generally bloomed. All the buds stored dry maintained their premature status irrespective of storage temperature. Storage of buds for 72 h at 5°C, followed by transferring to distilled water improved the longevity by about 5 to 6 days. Cold storage treatment before transferring to holding solution improved floral diameter, membrane integrity besides maintaining higher fresh and dry mass of flowers, sugar content, soluble proteins, and phenols. Our results suggested that wet and dry storage of premature buds of R. asiaticus for 72 h at 5°C, followed by placing them in distilled water, improved the cut flower longevity and can be used as effective postharvest storage treatments for this beautiful cut flower.     

低温预处理对甘蓝型油菜小孢子胚胎发生的影响

研究了大田环境下栽培的甘蓝型油菜小孢子胚状体诱导过程中低温预处理的作用,结果表明:两个供试材料经过低温预处理,小孢子胚胎发生能力明显提高,同时,不同基因型要求的预处理时间不同。G369的胚产量在低温3d时达到最高,是对照的27.2倍,而Topas的胚产量在低温2d时达到最高,是对照的15.4倍。试验还统计了子叶型胚状体发生频率,发现与对照相比,处理后子叶型胚状体的频率呈下降趋势,表明低温预处理使小孢子胚发育同步性降低。     

不同诱导培养基对小麦花药培养胚状体诱导率的影响

为明确W14-F诱导培养基对黑龙江省春小麦品种及人工合成小麦材料的花药培养胚状体诱导率,以6份人工合成小麦材料和4个普通小麦品种为材料,采用4种诱导培养基W14-F、MS、N6和C17进行花药培养比较试验。结果表明:不同的诱导培养基对于小麦材料的胚状体诱导率不同,4种诱导培养基的胚状体诱导能力依次为W14-F>C17>N6>MS,W14-F培养基胚状体诱导率明显高于其它3个培养基。人工合成小麦和普通小麦在4种培养基上都得到了相近的胚状体诱导率,说明W14-F培养基同样适合人工合成小麦。利用该技术体系培养人工合成小麦与普通小麦的杂种F1材料,得到了较高的诱导率,这为该技术体系直接应用于利用人工合成小麦改良小麦品种抗病抗逆性研究中提供了可靠的依据。     

花药培养建立辣椒DH纯系的初步研究

试验结果表明 ,基因型是影响辣椒花药培养胚状体诱导率的主要因素 ,激素是影响辣椒胚状体诱导率的重要因素 ;0 .1mg/L的 2 ,4-D和 0 .1mg/L的KT配比适合大部分辣椒基因型成功出胚 ,且出胚率较高。在不同的时期接种 ,相同基因型胚状体的诱导率存在显著差异。添加活性炭能提高胚状体的诱导率 ,在 0 .2 5 %～ 0 .5 %的浓度区间内得到的诱胚率差异不明显。     

碳源和植物生长调节剂对辣椒花药培养效率的影响（摘要）

[目的]为进一步提高辣椒花药培养效率。[方法]采用MS培养基,以博辣娇红为试材,研究碳源种类与浓度对花药愈伤组织诱导的影响;以佳玉为试材,按L16（4^5）正交试验设计,研究植物生长调节剂种类与浓度对辣椒花药培养效率的影响。[结果]愈伤组织和胚状体诱导率各浓度处理的均值麦芽糖虽略高于蔗糖,但未达差异显著水平;麦芽糖或蔗糖作碳源,3%～6%的浓度有利于提高愈伤组织与胚状体的诱导频率,超过6%以后,愈伤组织和胚状体诱导率随糖浓度的增加而极显著下降。生长调节剂对愈伤组织诱导率的影响为2,4-D﹥ZT﹥NAA﹥KT﹥6-BA;对胚状体诱导率的影响为2,4-D﹥NAA﹥ZT﹥KT﹥6-BA;2,4-D、ZT、NAA和KT对愈伤组织与胚状体诱导率均有显著或极显著影响;生长调节剂的使用浓度,2,4-D和ZT以1.0mg/L、NAA和KT对0.5mg/L的效果最好,ZT对提高愈伤组织和胚状体诱导率的效果比KT和6-BA都好;2,4-D1.0mg/L﹢ZT1.0mg/L﹢KT0.5mg/L﹢6-BA0.5mg/L是佳玉花药愈伤组织和胚状体诱导培养适宜的生长调节剂组合。[结论]该研究为辣椒花药培养研究提供了依据。     

Effects of Dietary Protein and Temperature on Growth and Flesh Quality of Songpu Mirror Carp

The study was conducted to demonstrate the effects of dietary protein and water temperature on growth and flesh quality of Songpu mirror carp （Cyprinus carpio） at an initial weight （165.24±5.08） g. Practical diets were formulated to contain five protein levels （29.12%, 31.46%, 34.49%, 38.17%, and 40.13%）, and each diet was randomly assigned triplicate groups of 15 fishes at three temperatures （18℃, 22℃, and 26℃） in the recirculation system. Fishes were fed twice daily to apparent satiation for 56 days. Results indicated that fishes had higher weight gain rate at 22℃ and 26℃ than that at 18℃ （P〈0.05）, but there were no significant differences between 22℃ and 26℃ （P〉0.05）. Based on the weight gain rate, dietary protein level 29.12% could meet the requirement of the body at 18℃, 22℃, or 26℃. Crude protein, crude lipid, moisture and ash of muscle had no significant differences among those treatments （P〉0.05）. pH （after 24 h） of muscle was the highest at 18℃ and the lowest at 22℃ （P〈0.05）, but no differences were observed among different protein level groups at each temperature （P〉0.05）. No significant differences on shear force, water holding capacity, collagen, glycogen and lactate among all the treatments were found （P〉0.05）. It was concluded that when C. carpio fed to apparent satiation, the growth mainly depended upon temperature. Dietary protein could not significantly affect flesh quality, but temperature significantly affected pH of muscles. A dietary protein level 29.12% could meet the requirement of the body at 18℃, 22℃, or 26℃.     

Expression of HSP72 in Mouse Preimplantation Embryos with Heat Shock

The objective of the paper was to detect HSP72 expression and HSP72 gene sequence in heat shocked mouse preimplantation embryos and the effects of different thermo conditions on hatching rates of embryos. The mouse blastocysts cultured in vitro were heat treated at 40℃ and 38℃ for 1 h, 2 h and 3 h and then recovered at 370C for 3 h, 2 h and 1 h, respectively, to detect their HSP72 gene expression by using RT-PCR after the total R.NA extraction. The hatching rate of the blastocysts for different treated groups was recorded and the expression of liSP72 in the blastocysts was determined by Western blot. The results showed that all the groups of blastocysts, including the control, had the expression of HSP72 gene. The expression of HSP72 protein had the highest level in the embryos stressed at 38℃ for 2 h, and it was significantly higher than that in the control group. The expression of HSP72 in the groups of blastocysts treated at 40℃ was not significantly different from that in the control group. The embryos with induction of mild heat shock at 38℃ for 2 h, then subjected to heat shock at 40℃ for 2 h, had a significant higher （P〈0.05） hatching rate of 54.74% compared to 47.85% in the embryos treated directly at 40℃ for 2 h. The above results indicated that the mouse blastocysts were sensitive to heat shock and a mild heat shock induced HSP72 gene expression. Induction of HSP72 expression with mild heat shock helped embryos to tolerate more severe heat shocks.     

Identification and characterization of a virus infecting pepper in Southern Primorskii Krai

Infection of pepper caused by an isometric virus with a 30 ± 3 nm virion diameter is revealed. The thermal inactivation point of virus in jimsonweed sap is 75–80°C, longevity in vitro 3 days at 25°C, and dilution end-point of virus 10^?3–10^?4. The capsid of the virus contains two types of protein with molecular mass 55 and 12.6 kDa. A distant antigenic relatedness to the tobacco ringspot virus of the genus Nepovirus is shown.     

萝卜游离小孢子培养及胚再生植株研究

[目的]探讨在自然生长条件下若干因素对萝卜（RaphanussativusL．）小孢子胚诱导及胚再生植株的影响。[方法]首先探讨不同萝卜品种的出胚能力,然后分析热激、低温预处理和培养基对小孢子培养的影响,最后研究胚状体再生成苗与植株继代培养情况。[结果]对22份萝卜材料进行游离小孢子培养,12份获得胚状体,3份获得再生植株,出胚材料中90％以上为杂种一代。4℃低温预处理0～3d小孢子出胚率无明显差异;培养基中加入1～2滴0．5％活性炭和一定浓度6-BA可显著促进小孢子诱导产生胚;不同萝卜品种适宜的热激温度和时间有所不同;发育正常的子叶型胚成苗率为100％;胚萌发分化早期适宜培养基为B5＋2％蔗糖＋1．2％琼脂,后期可逐渐将琼脂浓度降低;FCM测定小孢子植株自然加倍率为30％,单倍体占少数,四倍体与嵌合体（二、四倍）均为30％;秋水仙素处理单倍体植株,人工加倍率达60％。[结论]该研究完善了萝卜游离小孢子培养技术,可提高单倍体获得频率,加快这一技术在育种实践中的应用步伐。     

紫花苜蓿高频体胚发生及萌发成苗技术与转基因应用

以紫花苜蓿的子叶或叶片为外植体,建立高频体胚发生及成苗技术体系,为苜蓿耐逆性状的遗传改良提供技术支撑。在愈伤诱导、体胚诱导、体胚成熟与萌发等几个关键环节,分析比较培养基组分对培养效果的影响。建立在SHDK培养基上诱导愈伤、MB(-/+)+ABA 0.4mg/L+PEG-6000 50g/L+蔗糖50g/L培养基上诱导体胚、Bio2Y培养基上促体胚发育成熟、1/2 MS(或SH)培养基上体胚萌发成苗的高频再生技术体系。最适条件下,每克胚性愈伤组织可产生77.9个正常萌发成苗的健康体胚。再生植株在生长箱内经过2周20℃、16h/d的光照培养后移栽温室,成活率达90%以上。利用该离体再生技术成功地将凝集素基因PPA导入苜蓿中,获得抗蚜转基因苜蓿。     

Androgenic response of tomato (<Emphasis Type="Italic">Lycopersicon esculentum</Emphasis> Mill.) lines and their hybrids to anther culture

The androgenic ability of four tomato (Lycopersicon esculentum Mill.) lines (Micro-Tina, Micro-Gemma, Moneymaker and MsK8 as parents) and their hybrids (obtained as half diallel crosses) were studied through anther culture, The isolated anthers were cultured on three culture media including M1 (Park et al., 2001), M2 (Summers, 1997) and M3 (Evans et al. 1986) exposed to two pretreatments (26 ± 2°C as a control and at 4°C for 48 h as a cold pretreatment). Callus formation and callus growth (measured by callus diameter) occurred in the both pretreated anthers, but no callus produced shoot regeneration on the non-pretreated anthers. Statistical analysis showed significant differences between genotypes, culture media and their interactions for the studied traits. The highest callus formation was on M1 and M2 media, among which the longest callus diameter and the highest shoot regeneration were on M1 medium. The lines MsK8 and Micro-Gemma had a considerably higher frequency of callus formation and shoot regeneration and longer callus diameters than the other parents. On M1 medium, the highest frequency of callus formation and shoot regeneration belonged to hybrid MsK8 × Micro-Tina, and the longest callus diameter observed on hybrid Micro-Tina × Micro-Gemma. The mean values of callus formation, callus diameter, and shoot regeneration of hybrids in different culture media were significantly higher than those of the parents. In other words, heterosis (hybrid vigor) for the mentioned traits was observed on different culture media. Therefore, heterozygous genotypes (the hybrids) had a higher frequency of androgenic response than their parents.     

A promising alternate lipase for biodiesel fuel production

By screening samples from different sources, a high-yield yet low-cost lipase of Enterobacter aerogenes was found and used in biodiesel production. The enzyme activity of Enterobacter aerogenes reached 34.16 U·mL⁻¹ under the optimum conditions of initial pH 9.0, agitating at 30°C for 40 h, using NaHCO₃ and glucose as carbon sources and peptone as nitrogen source, with 0.1% (w/v) MgSO₄·7H₂O. When adding 1.0% (w/v) olive oil to the culture, the enzyme activity reached 66.31 U·mL⁻¹, which increased by 56.5% compared to that obtained by using 2.0% (w/v) olive oil as the sole carbon source. The lipase of Enterobacter aerogenes was immobilized by diatomite and used to catalyze biodiesel with rapeseed oil as raw materials, the transesterification rate was 92.91% under the optimum reaction conditions, which were using 1000 U immobilized lipase and 5 mL n-hexane, with ethanol as acyl acceptors, at the molar ratio of oil/ethanol of 1:4, and adding ethanol twice, at 35°C for 48 h. Therefore, the Enterobacter aerogenes will potentially serve as a promising alternative lipase for biodiesel production.     

Improvement of eggplant seed germination and seedling emergence at low temperature by seed priming with incorporation SA into KNO3 solution

The effects of incorporation SA into KNO₃ priming solution on the germination and emergence of eggplant seeds at 15°C were investigated. Seeds were primed into 3% KNO₃ containing 0.05, 0.1, 0.5, or 1mM salicylic acid (SA) for 6 days, respectively. After the priming, seeds were either immediately used for germination and emergence test at 15°C or stored at 4°C for 1 month and then for the germination test. The primed eggplant seeds in general improved the final germination percentage (FGP), germination rate (G₅₀), and germination synchrony (E_10–90) at 15°C compared with non-priming seeds. Priming seeds in 3% KNO₃ solution supplement with 0.1mM SA resulted in the best priming effect compared with other priming treatments and non-priming treatment, and all of the beneficial priming effects were still retained after stored at 4°C for 30 days. These results indicated that priming seeds in 3% KNO₃ solution containing 0.1mM SA could be used as an effective method to improve low-temperature performance of eggplant seeds and subsequent seedling growth.