金星无核葡萄种胚培养最佳接种期的选择

金星无核葡萄的胚珠可正常受精，绝大多数受精胚在花后21d开始败育。当胚发育最为充分而又尚未败育时取胚珠进行培养，存活胚的比例较高。金星无核葡萄最适宜的接种时期为花后36～39d，此时正是浆果开始软化和着色前期。     

'森田尼无核'葡萄胚发育及败育的细胞学特征

以种子败育型葡萄‘森田尼无核’为试材,系统研究其胚发育及败育的细胞学进程,分析胚乳败育与胚败育的关系。结果表明:种子败育型葡萄‘森田尼无核’与有核葡萄‘谢花红’的胚均能发育到球形胚阶段,随后‘森田尼无核’的胚逐渐败育,而‘谢花红’的胚珠继续发育为成熟种子。‘森田尼无核’的胚乳败育早于胚败育,其胚挽救适宜取材时期是花后28 d。     

无核葡萄胚败育生理生化因子灰色关联分析

以无核葡萄‘无核翠宝’、‘丽红宝’和有核葡萄‘赤霞珠’(对照)为试材,观测其发育过程中果实和胚珠的形态学、生理学变化,利用主成分分析法对6个生理生化指标进行分析,结合灰色关联度分析,筛选出与无核葡萄胚败育相关的主要生理生化因子。结果表明,‘无核翠宝’和‘丽红宝’分别于花后36 d和32 d胚珠畸形率达最高,开始败育。随着果实发育,3个葡萄品种胚珠中丙二醛(MDA)含量呈上升趋势;果肉中可溶性糖和可溶性蛋白含量均呈上升趋势,但在无核葡萄胚珠中先增后降,在败育时期达到最大;POD和CAT活性在两个无核品种中先升后降,在败育时期的胚珠中显著高于有核葡萄,而在果肉中显著低于有核葡萄;胚珠的SOD活性在无核葡萄败育前显著增加,在有核葡萄中则呈上升趋势。综合分析,无核葡萄果肉中可溶性糖、可溶性蛋白含量和胚珠中MDA、可溶性蛋白含量,SOD活性等生理指标为无核葡萄胚败育的主要影响因子,其中胚珠SOD活性与无核葡萄胚败育关联度最高,其次是果肉可溶性蛋白含量和胚珠数。本研究为今后无核葡萄胚挽救育种采样时期的确定提供了理论指导。     

“新葡7号”葡萄胚挽救最佳接种时期研究

正近年来,随着生物技术的发展,胚挽救技术在葡萄育种上的研究应用,为培育优质无核葡萄品种提供了新思路和崭新的途径,丰富了无核葡萄有性杂交的组合方式,提高了无核葡萄品种的选育效果。无核葡萄胚的发育受基因型的严格控制。胚龄越小越难培养,而胚龄越大则存在胚萎缩退化的可能,所以掌握无核葡萄胚珠败育时期,确定胚珠接种的适宜时间,是胚珠培养能否成功的前提。郭修武等人研究了胚珠接种时间和培养基种类对无核葡萄胚挽救的影     

外源6-BA对汤姆逊无核葡萄胚珠败育及胚培养的影响

以汤姆逊无核葡萄为试材,研究不同浓度6-BA对葡萄胚珠败育及胚培养的影响。结果表明:花前2周用不同浓度的6-BA处理汤姆逊无核葡萄花穗,可以延缓胚珠败育5 d以上;6-BA处理的胚珠在花后45 d接种萌发率达到最高,均在6.9%以上,是对照的1.7倍。说明6-BA可以延缓胚珠败育,扩大无核葡萄育种母本选择范围,提高无核葡萄育种效率。     

无核葡萄与中国野生葡萄杂种胚败育的某些生理生化变化

 通过对无核葡萄、有核葡萄以及无核葡萄与中国野生葡萄杂种的胚珠进行离体培养, 并结合生理生化指标测定发现: 不同的无核葡萄及其杂种胚败育的时间不同, 胚败育时, 保护酶活性下降, 反应膜脂过氧化水平的MDA 含量上升, 可溶性糖与可溶性蛋白质含量降低, 可溶性淀粉含量变化不大。进行胚挽救时, 应根据不同的杂交组合, 在最佳时间, 即胚发育程度最高、MDA 含量较低时取胚进行培养。     

‘杨格尔’与‘京秀’葡萄胚珠发育过程中活性氧代谢的比较研究

以盛花后不同时期的无核葡萄品种杨格尔和有核葡萄品种京秀的胚珠为材料,测定胚珠中超氧阴离子和过氧化氢（H₂O₂）含量及活性氧清除酶活性的变化。结果表明：在两品种胚珠发育过程中均呈一直上升的趋势,在胚珠发育的中后期无核品种扬格尔产生速率明显大于有核品种京秀;胚珠中H2O2含量表现先升后降再升的趋势,但再升时无核品种扬格尔显著大于有核品种京秀;胚珠内活性氧清除酶超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、 抗坏血酸过氧化物酶（ASA-POD） 及无核品种扬格尔的过氧化物酶（POD）表现先升后降的趋势,无核品种扬格尔的酶活性在胚珠败育后明显下降,有核品种京秀仍在较高的水平,且有核品种的POD活性先快速上升后一直保持缓慢上升的趋势。胚珠内活性氧类物质及活性氧清除酶类的变化趋势与无核葡萄杨格尔幼胚败育有一定相关。     

''''杨格尔''''与''''京秀''''葡萄胚珠发育过程中的活性氧代谢比较

以盛花后不同时期的无核葡萄品种杨格尔和有核葡萄品种京秀的胚珠为材料,测定胚珠中超氧阴离子(O2.-)生成速率和过氧化氢(H2O2)含量及活性氧清除酶活性的变化。结果表明:O2.-在两品种胚珠发育过程中均呈一直上升的趋势,在胚珠发育的中后期无核品种杨格尔产生速率明显大于有核品种京秀;胚珠中H2O2含量表现先升后降再升的趋势,但再升时无核品种杨格尔显著大于有核品种京秀;胚珠内活性氧清除酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(AsA-POD)及无核品种杨格尔的过氧化物酶(POD)活性表现先升后降的趋势,无核品种杨格尔的酶活性在胚珠败育后明显下降,有核品种京秀仍在较高的水平,且有核品种的POD活性先快速上升后一直保持缓慢上升的趋势。胚珠内活性氧类物质及活性氧清除酶类的变化趋势与无核葡萄杨格尔幼胚败育有一定相关。     

种子败育型葡萄胚珠中内源多胺含量与胚珠发育及败育的关系

为探明种子败育型葡萄胚珠中内源多胺含量与胚败育的关系,以种子败育型葡萄和有核对照品种自交果实为试材,研究了不同胚胎发育时期胚珠内源多胺含量的变化规律。结果表明,除有核葡萄Spm含量在胚发育过程中始终表现上升趋势外,葡萄胚珠内源PAs、Put和Spd含量均表现为先升后降的变化趋势。种子败育型葡萄的内源PAs、Put和Sp...     

假单性结实型无核葡萄胚挽救影响因子研究

【目的】探究无核葡萄胚发育和胚萌发的影响因子,有利于优化无核葡萄胚挽救技术体系及提高其育种效率。【方法】以早黑宝盛花后100 d种胚为试材,采用剥种取胚进行胚离体培养,研究外源激素和营养添加物质对胚萌发成苗的影响,基于熵权TOPSIS(优劣解距离法)分析法筛选出最佳胚萌发培养基;以无核翠宝(盛花后28～38 d)、丽红宝(盛花后28～32 d)和晶红宝(盛花后28～32 d)未成熟幼果为试材,研究不同取样时间和不同培养时间对胚珠发育率的影响,基于熵权TOPSIS分析法综合评价不同形态胚的萌发成苗效果。【结果】在胚萌发培养基中添加0.1 mg·L-1IAA+0.2 mg·L-16-BA+0.5 g·L-1葡萄汁时,胚的萌发率和成苗率显著高于其他。取样时间接近(或达到)胚败育始期时,胚发育率最高。无核翠宝和丽红宝胚珠离体培养10周后的胚萌发率最高(≥50.00%)。不同形态胚的萌发成苗效果不同,子叶形胚的萌发成苗效果最好,其他排序为:鱼雷形胚﹥球形胚﹥心形胚﹥畸形胚。【结论】胚萌发培养基中添加外源物质有利于胚萌发成苗;离体培养败育时期的胚珠能获得更高的胚发育率和多胚率;子叶形胚和鱼雷形胚更易萌发成苗。     

早熟和特早熟桃胚珠培养研究

本文以自然授粉的‘春蕾’、‘沪017’、‘布目早生’、‘1-4-8’、‘7-6-9’的胚珠为试材,研究了早熟和特早熟桃胚珠培养胚成苗的有关影响因素。本试验5个品种(品系)的结果表明,桃品种或品系的果实生育期愈短,胚珠培养胚的成苗率愈高,特早熟桃‘春蕾’花后48天的胚珠培养在改良S H培养基附加BA1.0mg/1 IAA1.0mg/1上30天,将长大的幼胚转移至TM培养基上,胚的成苗率达75％。此外,作者对胚珠培养技术在桃育种中的应用范围及其培养程序开展了讨论。     

Consequences to fertilization of the developmental stage of apricot ovules at anthesis

Summary

Flowers at anthesis from the apricot cvs Palstein and Goldrich were examined to determine their ovule maturity stage. Further, flowers from both cultivars were pollinated and collected after 2000 and 3000 growing degree hours (GDH) in two consecutive years. Numbers of fertilized ovules were recorded in those flowers. Ovules of ‘Palstein’ from flowers at anthesis showed predominantly an embryo sac at the eight nuclei stage of development while embryo sacs in ‘Goldrich’ were mainly in a four nuclei stage. After 2000 GDH from pollination 90% of viable ovules were fertilized in ‘Palstein’ whereas only 37.5% of them were fertilized in ‘Goldrich’. After 3000 GDH, Goldrich presented the same percentage of fertilized ovules that was recorded after 2000 GDH. Non-fertilized ovules did not degenerate and developed normally in pollinated and non-pollinated flowers from both cultivars. Further, non-fertilized pollinated flowers were not different from non-pollinated flowers in the development of the embryo sac, indicating that the chances for fertilization would not be increased by pollination in apricot. We suggest that the lack of fertilization in ‘Goldrich’ is due to degeneration of the male gametophyte, which would have a limited life, not long enough for the embryo sacs to develop to maturity.     

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.     

Ovules that failed to form seeds in zinnia (Zinnia violacea Cav.)

The structure of ovules that failed to form seeds after compatible pollination in double-flowered zinnias (Zinnia violacea Cav.) was investigated by observing histological sections, and comparing the normal and abnormal development of the seeds. In an embryo sac of zinnia at anthesis, a large nucleus of fused polar nuclei was clearly recognized. Success in double fertilization was determined by the disappearance of this nucleus. In normally developing seeds, the cellular endosperm became vague at 5–6 days after pollination (DAP) and the developing embryos occupied the entire portion of the seeds up to 10 DAP. A number of ovules failed in fertilization for the lack of an embryo sac. In most of the aborted seeds, the embryo sacs degenerated, whereas the aborted embryos were still alive in a small number of aborted seeds. A small percentage of the aborted seeds exhibited an aborted embryo and an unfertilized fused nucleus of two polar nuclei or ‘single fertilization’. The three major problems suggested for the failure in seed formation in pollinated florets of zinnia included failure in fertilization, ovules lacking embryo sacs, and abortion of developing seeds.     

离体葡萄未成熟胚成苗途径研究

以巨峰葡萄剥离幼胚及红井川品种切喙胚珠为外值体接种至含不同浓度２，４Ｄ、ＮＡＡ与６－ＢＡ配比的ＮＮ－１９６９培养基中，诱导葡萄未成熟胚产生胚状体及实现胚萌发。两品种均是在含２，４－Ｄ０．５ｍｇ·１－１、６－ＢＡ１．０ｍｇ·１－１的培养基中诱导胚进入胚状体发生途径的；而在含ＮＡＡ（１．０、２．０ ｍｇ·１－１）与 ６－ＢＡ（０．２、１．０）或２．０ ｍｇ·１－１ＺＴ 配比的培养基上幼胚进入胚萌发途径成苗。由此讨论了培养基激素、胚发育时期对胚培养两种途径的诱导和控制。     

黄瓜未受精胚珠离体培养及单倍体植株再生

以欧亚杂交类型黄瓜（Cucumis sativus L.）65G × 228 的F1 为材料,对未受精胚珠进行离体培养,通过6 因素5 水平的正交试验,研究预处理温度、诱导培养基、TDZ、KT 和BA 浓度对未受精胚珠雌核发育的影响。结果证明TDZ 浓度对胚发生频率有极显著影响,是诱导黄瓜离体雌核发育的最主要因素,当其浓度为0.02 mg · L-1 时胚状体的诱导率最高。诱导培养基和KT 浓度显著影响胚发生率,而预处理温度和BA 浓度影响不显著。硝态氮（NO3-）和铵态氮（NH4+）含量比例均接近1∶1 的诱导培养基,较其他比值较大的培养基的胚发生率有显著提高。利用流式细胞仪进行倍性鉴定,再生植株中单倍体率为18.2%。     

红宝石无核葡萄胚珠培养成苗技术研究

以自然授粉的红宝石无核为试材,观察了胚珠生长状况,对胚珠离体培养与成苗技术进行了探索,结果表 明:红宝石无核的胚珠发育在花后35 d是其胚挽救的最佳时期。此时,取出胚珠接种在Nitsch+IAA 2.0 mg/L+GA 0.4 mg/L,附加水解酪蛋白100mg/L,活性炭2g/L的培养基中培养120 d,胚发育率为58％,后将胚珠切开处理并转接 到胚萌发培养基1/2MS+6-BA 0.5 mg/L中萌发,萌发率为16％,在1/2MS+6-IBA 0.2mg/L培养基上成苗,小苗生长 最好。通过前期保湿等措施,小苗移栽成活率达100％。     

杏胚珠培养及幼胚子叶不定芽诱导研究

以盛花后25、35、50、55、60d的凯特杏胚珠为外植体,研究胚珠培养的适宜条件;选择发育一致的幼胚子叶,研究不定芽的发生条件。结果表明,SH培养基是凯特杏胚珠培养的适宜培养基;盛花后25d的凯特杏胚珠(PF=0),仅采用生长培养不足以发育成可见胚,试验采用诱导方式获得了可见胚,诱导出胚率为80%;以MS为基本培养基,BA与NAA结合诱导能力强;但这种胚小且瘦弱,继续生长培养,利用幼胚子叶诱导不定芽。在幼胚子叶诱导不定芽方面,盛花后25、35d取样的凯特杏胚珠,培养后获得幼胚子叶,诱导不定芽的适宜TDZ质量浓度为1.25mg/L,诱导出芽率分别为85.00%、80.00%;盛花后50、55d取样的材料,适宜TDZ质量浓度为2.50mg/L,出芽率分别为65.00%、60.00%,而盛花后60d取样的材料,适宜TDZ质量浓度为5.0mg/L,出芽率为61.11%,不定芽主要发生在子叶的近轴面、胚轴连接处及子叶近轴端两侧;在相同培养基中,25℃进行21d暗培养,盛花后35、50、55、60d材料出芽率分别为72.22%、65.00%、60.00%、55.56%;4℃下28d暗培养,35、50、55、60d依次为59.09%,71.42%、77.78%、86.36%;对不定芽进行伸长、生根与快繁培养,获得了健康正常的植株。通过以上研究,建立了凯特杏幼胚4级培养体系,即(1)胚珠培养,获得可见胚(1级培养);(2)幼胚生长培养(胚培养),获得了健壮幼胚(2级培养);(3)幼胚子叶再生,获得不定芽(3级培养);(4)不定芽组培苗快繁(4级培养)。     

On the senescence of ovules in cherries

The longevity of ovules in sweet and sour cherries has been studied in vivo, using a fluorescence—microscopic method which enables one to determine the viability of ovules at a very early stage. It was found that ovules which were not fertilized became progressively senescent 4 to 5 days after anthesis, and about a week after flower opening a rather high percentage of the ovules was non-viable.Applied growth regulators usually enhanced senescence of the ovules, especially GA₃. Even those that delayed senescence of the external flower parts, e.g. 2,4-D, benzyladenine and kinetin, accelerated the ageing of ovules in most instances. No substance was found which extended the longevity of ovules in cherries.     

Growth and development of ovules of banana,plantain and enset (Musaceae)

Sterility has profound effects on the development of reproductive tissues in members of the Musaceae and limits genetic improvement required to deal with new diseases. Ovules of seeded diploid Musa acuminata (AA) and edible triploids (AAA) from the same cytogenetic group, plus edible triploids containing genomes of the M. balbisiana cytogenetic group (AAB, ABB) and the related genus Ensete sp. were studied to determine the effects of sterility on the growth and development of the ovule and its tissues. Specimens were collected from subtropical and Mediterranean environments in Australia.At anthesis, the ovules of triploid plants were 36% larger than the ovules of diploid plants. The diploid ovules ceased growth shortly after the inflorescence emerged from the pseudostem. In contrast, the triploid ovules continued to grow 7–10 days past anthesis, increasing in size by 70%.All ovules of diploid M. acuminata ssp. had an embryo sac at anthesis, against 97% for triploids. At anthesis the embryo sacs in diploid ovules occupied 2.7% of the nucellus compared with 1.5% in triploid ovules. The embryo sacs did not grow between bunch emergence and anthesis, once formed they maintained the same size. Many embryo sacs were not positioned correctly, flush with the nucellar cap. The diploid M. acuminata ssp. had 75% of embryo sacs correctly positioned against 10% in the edible triploids. The proportion of balbisiana genome (B) did not affect ovule or nucellus size or shape, or cell number across the nucellus. It increased the embryo sac presence 96–100% of ovules.The sterile edible triploid bananas have embryo sacs at anthesis but many are incorrectly positioned, which may contribute to their sterility. The balbisiana genome in the edible triploids was associated with a 2.4-fold increase in the number of correctly positioned embryo sacs and this may contribute to the increased fertility associated with the B genome.