荔枝和龙眼种子胚轴的脱水及贮存

新采收的荔枝和龙眼种子的离体胚轴,在pH4．8—8．0的WPM培养基中均可正常生长成苗,加入KTt和NAA对胚轴的生长有一定的抑制作用。荔枝和龙眼种子离体胚轴的临界含水量分别为30％和23%,低于此阈值时,生活力迅速下降,且培养25天不能成苗。经短期贮存的荔枝和龙眼种子离体旺轴生活力下降很快,其中脱水的较未脱水的旺轴下降快,且贮存后的旺轴接种后易感染。脱水至临界古水量的龙眼离体胚轴经液氟贮存后约有25.50％的存活率,荔枝胚轴则不能存活。     

荔枝和龙眼种子发育过程中ABA含量及对外源ABA敏感性的变化

在荔枝和龙眼种子发育过程中,内源ＡＢＡ水平先是上升,至大约７８～８０ＤＰＡ时出现高峰,之后两者ＡＢＡ含量均不断下降。果实成熟时采收的种子,ＡＢＡ含量比高峰时分别下降近６倍。另外,随着种子的发育,种子及其胚轴对外源ＡＢＡ的敏感性（ＳＡＢＡ）亦持续下降。１０－４ｍｏｌ／ＬＡＢＡ可以完全抑制９０ＤＰＡ前的荔枝和龙眼种子的萌发,但对成熟种子１０－２ｍｏｌ／ＬＡＢＡ亦不能抑制其萌发。龙眼种子离体胚轴的ＳＡＢＡ高于荔枝。ＡＢＡ含量与敏感性的这种变化可能是两种顽拗性种子成熟时萌动,进而不耐脱水贮藏的重要原因之一。     

荔枝和龙眼种子发育过程中ABA含量及对源ABA敏感性的变化

在荔枝和龙眼种子发育过程中,内源ＡＢＡ水平先是上升,至大约７８－８０ＤＰＡ时出现高峰,之后两者ＡＢＡ含量均不断下降。果实成熟时采收的种子,ＡＢＡ含量比高峰时分别下降近６倍。另外,随着种子的发育,种子及其胚轴对外源ＡＢＡ的敏感性亦持续下降,１０＾－４ｍｏｌ／ＬＡＢＡ可以完全抑制９０ＤＰＡ前的荔枝和龙眼种子的萌发,但对成熟种子１０＾－２ｍｏｌ／ＬＡＢＡ亦不能抑制萌发。龙眼种子离体胚轴的ＳＡＢＡ高于荔枝     

木波罗种子脱水敏感性与膜脂过氧化的研究

刚采收的木波罗种子含水量为５８．６％。随着含水量下降,种子的发芽率和发芽指数迅速下降,种子对脱水非常敏感,是典型的顽拗性种。自然脱水时,种子胚轴和子叶中超氧物歧化酶的活性先上升,然后下降,丙二醛和脂质氢过氧化物的含量显著增加。其脱水敏感性的原因可能是当种子脱水时,植物酶ＳＯＤ的活性下降,膜脂过氧化作用加强,从而使膜的结构和功能受到破坏,种子生活力丧失。     

顽拗性竹柏种子的贮藏特性?

文章对竹柏( Podocarpus nagi)种子的脱水耐性和贮藏特性进行了研究,结果表明：竹柏种子成熟时初始含水量约为(35±0?7)％,种子对脱水敏感,其最低安全含水量约为(16?86±0?73)％,具有顽拗性种子的典型特征;湿藏和半干藏都可以作为短期保存竹柏种子的方法,且以4℃保存优于15℃保存,但不管种子含水量如何,零下低温保存对竹柏种子都是致命的;半干藏法保存实验中,未进行脱水处理的种子(对照)在4℃贮藏6个月,种子萌发率没有发生明显下降,但贮藏期延长到9个月时,临界含水量的种子萌发力保存最高;不管贮藏介质的含水量高低,也无论贮藏在4℃还是15℃,湿藏种子在9个月的贮藏期内萌发率均没有明显的降低,但当贮藏到12个月时,15℃湿藏种子的萌发率显著高于4℃贮藏的种子,但15℃湿藏的种子在贮藏到3个月时即发现种子在贮藏期间萌发,且随着贮藏介质含水量的升高和贮藏期的延长,萌发的种子增多;竹柏的离体胚经过2h硅胶快速脱水至含水量7％后再冷冻即可获得90％以上的融后存活率,且超低温保存1年的离体胚解冻后,与只保存1周的存活率没有明显差异,表明超低温长期保存竹柏种子是可行的。本研究可以为进一步探究顽拗性种子的短期贮藏和长期保存提供理论基础和基础资料。     

分子运动性预测麻栎种子离体胚轴适宜贮藏条件初探

应用电子顺磁共振波谱仪和自旋标记技术,以硝基氧探针为标记物,检测了室温下麻栎种子离体胚轴脱水过程中分子运动性的变化。发现含水量0.7gH2O/gDW至0.64gH2O/gDW范围是细胞质粘度的转折区域,低于这个含水量区域,细胞质粘度骤然上升,推测这个区域是室温下保存离体胚轴的适宜含水量下限。通过变温电子顺磁共振波谱测定,找到离体胚轴含水量在0.43gH2O/gDW至1.02gH2O/gDW范围内,分子运动性的临界温度和玻璃态相变温度所在区间。根据分子运动性随温度变化的规律,预测含水量为0.69gH2O/gDW的麻栎种子离体胚轴适宜贮藏温度约为-50℃。     

快速干燥对黄皮胚轴贮藏寿命及抗氧化酶活性的影响

以黄皮种子离体胚轴为材料 ,研究了快速干燥对胚轴贮藏寿命的影响及与抗氧化酶活性的关系。未经脱水的胚轴在 15℃贮藏 12 8d仍具有较高的存活率和活力指数。快速脱水 1.5和 3.0h后胚轴的存活率和活力指数并未因脱水而下降 ,但它们在 15℃贮藏的寿命分别只有 8和 2d。未经脱水胚轴的抗氧化酶(SOD、POD、CAT)活性在贮藏过程中能较好地保持 ,但经快速脱水后其抗氧化酶活性在贮藏过程中则迅速下降 ,脱水时间越长 ,酶活性在贮藏过程中的下降就越快。     

脱水速率对黄皮胚轴脱水敏感性及膜膜过氧化的影响

以黄皮种子离体胚轴为材料,研究了不同于燥速率对胚轴脱水反应和膜脂过氧化的影响。在脱水过程中,胚轴的萌发率,活力指数,电解质渗漏速率,超氧化物歧化酶（SOD）,过氧化物酶（POD）和过氧化氢酶（CAT）活性逐渐降低,膜脂过氧化产物MDA的含量不断增加,脱水速率愈快,胚轴的半致死含水量就愈低。快速干燥的胚轴能在较低的含水量存活是因为顾中间含量下发生的膜脂过氧化作用的时间,以及保持较高的SOD,POD和CAT活性,缓慢干燥的胚轴当与周围环境达到水分平衡后,生活力的丧失将与保持在水分平衡后的时间有关,。因此,脱水速率是一种影响顽拗性种子或者胚轴脱水敏感性的重要因子。     

脱水速率对黄皮胚轴脱水敏感性及膜脂过氧化的影响

以黄皮种子离体胚轴为材料,研究了不同干燥速率对胚轴脱水反应和膜脂过氧化的影响.在脱水过程中,胚轴的萌发率、活力指数、电解质渗漏速率,超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性逐渐降低,膜脂过氧化产物MDA的含量不断增加.脱水速率愈快,胚轴的半致死含水量就愈低.快速干燥的胚轴能在较低的含水量下存活是因为缩短了在中间含水量下发生的膜脂过氧化作用的时间,以及保持较高的SOD、POD和CAT活性;缓慢干燥的胚轴当与周围环境达到水分平衡后,生活力的丧失将与保持在水分平衡后的时间有关.因此,脱水速率是一种影响顽拗性种子或者胚轴脱水敏感性的重要因子.     

顽拗性竹柏种子的贮藏特性

文章对竹柏(Podocarpus nagi)种子的脱水耐性和贮藏特性进行了研究,结果表明:竹柏种子成熟时初始含水量约为(35±0.7)%,种子对脱水敏感,其最低安全含水量约为(16.86±0.73)%,具有顽拗性种子的典型特征;湿藏和半干藏都可以作为短期保存竹柏种子的方法,且以4℃保存优于15℃保存,但不管种子含水量如何,零下低温保存对竹柏种子都是致命的;半干藏法保存实验中,未进行脱水处理的种子(对照)在4℃贮藏6个月,种子萌发率没有发生明显下降,但贮藏期延长到9个月时,临界含水量的种子萌发力保存最高;不管贮藏介质的含水量高低,也无论贮藏在4℃还是15℃,湿藏种子在9个月的贮藏期内萌发率均没有明显的降低,但当贮藏到12个月时,15℃湿藏种子的萌发率显著高于4℃贮藏的种子,但15℃湿藏的种子在贮藏到3个月时即发现种子在贮藏期间萌发,且随着贮藏介质含水量的升高和贮藏期的延长,萌发的种子增多;竹柏的离体胚经过2 h硅胶快速脱水至含水量7%后再冷冻即可获得90%以上的融后存活率,且超低温保存1年的离体胚解冻后,与只保存1周的存活率没有明显差异,表明超低温长期保存竹柏种子是可行的。本研究可以为进一步探究顽拗性种子的短期贮藏和长期保存提供理论基础和基础资料。     

逐步提高培养基蔗糖浓度诱导的黄皮胚轴耐脱水性与细胞超微结构的变化

黄皮胚轴在蔗糖浓度按27%→50%→60%递增的WPM培养基中培养后耐脱水性显著提高,大部分胚轴脱水至含水量为18.8%时仍具有再生植株的能力.超微结构观察表明,对照下胚轴的细胞在脱水至36.3%含水量时质膜和液泡解体,叶绿体和线粒体崩溃.而经蔗糖逐步加浓培养的胚轴脱水至35.2%含水量时,大部分细胞发生质壁分离,细胞质稠密,叶绿体内部的淀粉粒变大;脱水至18.8%时细胞质壁分离加剧,大部分叶绿体和线粒体局部损伤.脱水至18.8%的胚轴重新吸胀4 d后细胞损伤被修复.     

In vitro propagation of Allium tuberosum Rottl. ex. Spreng. by shoot proliferation

Seeds of trifoliate orange (Poncirus trifoliata (L.) Raf.) are sensitive to desiccation, and could not withstand reduction in moisture level below 20%, whereas the excised embryonic axes could be easily desiccated to moisture levels as low as 14% without much loss in viability. Axes could be successfully cryopreserved in liquid nitrogen (–196°C) for eight months. The viable embryonic axes exhibited good growth on modified Murashige and Skoog medium supplemented with 1-Naphthalene acetic acid (NAA) and 6-Benzylaminopurine (BAP). Growth of cryopreserved axes was promoted in the presence of charcoal in the medium allowing for plant recovery.Abbreviations NAA Napthaleneacetic acid - BAP 6-Benzylamino-purine - MS Murashige and Skoog (1962) - LN Liquid nitrogen     

Cryopreservation of embryonic axes of trifoliate orange (Poncirus trifoliata [L.] RAF.)

Seeds of trifoliate orange (Poncirus trifoliata (L.) Raf.) are sensitive to desiccation, and could not withstand reduction in moisture level below 20%, whereas the excised embryonic axes could be easily desiccated to moisture levels as low as 14% without much loss in viability. Axes could be successfully cryopreserved in liquid nitrogen (–196°C) for eight months. The viable embryonic axes exhibited good growth on modified Murashige and Skoog medium supplemented wiith 1-Naphthalene acetic acid (NAA) and 6-Benzylaminopurine (BAP). Growth of cryopreserved axes was promoted in the presence of charcoal in the medium allowing for plant recovery.Abbreviations NAA Napthaleneacetic acid - BAP 6-Benzylaminopurine - MS Murashige and Skoog (1962) - LN Liquid nitrogen     

Ascorbic acid metabolism in ageing recalcitrant sal (Shorea robusta Gaertn. f.) seeds

Changes in ascorbate content and its enzymatic utilization pattern were studied in embryonic axes and cotyledons of sal seeds undergoing rapid loss of viability, at ambient conditions. Ascorbate levels were significantly higher initially in the embryonic axes (0.32 mg/g fresh weight) and cotyledons (0.21 mg/g fresh weight) of freshly mature, relatively hydrated (42.2% moisture content) and 100% viable sal seeds. It declined sharply as the tissues; embryonic axes and cotyledons, desiccated with absolutely no detectable amount in non-viable seeds (21% moisture content). Significantly strong correlation was obtained between desiccation of embryonic axes (r = 0.96) and cotyledon (r = 0.97) with loss of ascorbate levels and loss of germinability. Higher rates of ascorbic acid utilization (AAU) recorded in the embryonic axes of 100% viable seed declined sharply as the seed viability reduced due to desiccation below 36.8% moisture content. AAU was not detected in the cotyledons.     

Cryopreservation of Quercus faginea embryonic axes

Embryonic axes, excised or included in cotyledonary tissue, of Quercus faginea were tested for viability after rapid freezing and storage in liquid nitrogen (−196 °C). Explants were previously pretreated by desiccation at different moisture contents or by soaking in cryoprotectant (15% dimethyl sulfoxide). Best germination response after freezing (60%) was observed when embryonic axes were desiccated from 53 to 21% moisture content (on a fresh weight basis). Desiccation below 39% moisture content or freezing produced damage resulting in loss of organized growth and the induction of callus formation.     

Critical moisture content for successful cryopreservation of embryonic axes of <Emphasis Type="Italic">Fortunella polyandra</Emphasis> determined by differential scanning calorimetry (DSC)

The relationships between water content of desiccated embryonic axes (using different methods of desiccation), the availability of water determined by differential scanning calorimetry (DSC) analysis and recovery percentage after liquid nitrogen (LN) exposure of Fortunella polyandra embryonic axes were investigated. The objectives were to understand thermal properties of desiccated embryonic axes during cryopreservation and to determine the critical moisture contents for successful cryopreservation of the embryonic axes. Excised embryonic axes were desiccated under laminar air flow (0, 10, 15, 30 and 45 min), over silica gel (0, 5, 15, 30 and 60 min), and ultra-rapidly (0, 5, 10, 20 and 25 min). Desiccation under laminar air flow resulted in an optimal water content of 0.150 gH₂O g^?1 dw and a survival of 50 % after cryopreservation, while the unfrozen water content (WC_u) was 0.126 gH₂O g^?1 dw. After drying over silica gel, the optimal water content was 0.190 gH₂O g^?1 dw, where the survival was 40 % after cryopreservation and the WC_u was determined as 0.177 gH₂O g^?1 dw. Using the flash-drying method, the optimal water content was found to be 0.145 gH₂O g^?1 dw, the survival was 50 % after cryopreservation and the WC_u was 0.133 gH₂O g^?1 dw. Embryonic axes of F. polyandra showed low-to-moderate tolerance to desiccation. The results of the freezing transitions for all the desiccation times and methods showed that the onset temperature and the peak of the mean enthalpy decreased in size with decreasing water content. DSC elucidated the critical moisture contents and the cooling and melt enthalpies for successful cryopreservation of F. polyandra embryonic axes.     

Desiccation-induced changes in lipid peroxidation, superoxide level and antioxidant enzymes activity in neem (Azadirachta indica A. Juss) seeds

The freshly harvested mature neem seeds (42.2 % seed moisture content) with 100 % viability deteriorate when naturally desiccated to below 10.9 %. The desiccation-induced loss of viability was closely associated with over accumulation of superoxide anion and lipid peroxidation products both in the embryonic axes and cotyledons. The levels of superoxide anion and lipid peroxidation products were higher in axes compared to cotyledons. Superoxide dismutase activity was not much affected, both in the axes and cotyledons of 100 % viable seeds during desiccation from 42.2 % to 10.9 % seed moisture content. Steep rise in its activity was observed during drying below lowest safe moisture content (LSMC). Activities of catalase and peroxidase exhibited substantially higher levels in the 100 % viable seeds dehydrated up to LSMC. Their activities declined sharply in seeds with water content below LSMC. Impairment of catalase and peroxidase activities possibly lead to enhanced accumulation of reactive oxygen species. The accumulation of superoxide anion, lipid peroxidation and differential expression of superoxide dismutase and catalse/peroxidase activities in response to desiccation (below LSMC) is discussed to explain the intermediate storage physiology of neem seeds.     

The protective role of selenium in recalcitrant Acer saccharium L. seeds subjected to desiccation

Freshly harvested silver maple (Acer saccharinum L.) seeds were soaked in either sodium selenite (10 mg/L) or water for 6 h. After washing and air drying, seeds were desiccated at 22 °C at a RH of 45-50% to comparable water levels from 50 to 12%. Germination capacity was significantly higher in seeds treated with selenium and desiccated [from 50 to 40, 35 and 30% of water content (WC)] than in water-soaked seeds. At 20% WC, the seeds from both treatments had low viability (approximately 20%). The electrolyte leakage and the MDA content were significantly lower in the embryonic axes of seeds soaked in selenite than in seeds soaked in water. We also found that the activity of glutathione peroxidase (GPX) of embryonic axes from selenium-treated seeds that were not desiccated, or from seeds that were desiccated to 40 and 35% WC, was significantly higher than that of non-treated axes. No difference in GPX activity was detected in cotyledons. This was confirmed by activity staining of GPX after native PAGE of proteins extracted from embryonic axes and cotyledons. An increase in glutathione reductase (GR) activity was also observed in embryonic axes of seeds treated with selenium and dried to 35 and 30% WC compared to non-treated samples. Selenium appeared to have no such effect on cotyledons.     

Cryopreservation of protocorm-like bodies (PLBs) of <Emphasis Type="Italic">Phalaenopsis bellina</Emphasis> (Rchb.f.) christenson by encapsulation-dehydration

Protocorm-like bodies (PLBs) of Phalaenopsis bellina were successfully cryopreserved by the encapsulation-dehydration approach. Various stages in obtaining successful cryopreservation using this method were optimized. Encapsulated PLBs precultured in half-strength MS medium supplemented with 0.75 M sucrose for 3 days exhibited the highest viability in terms of 2,3,5-triphenyltetrazoliumchloride (TTC) reduction. The amount of sucrose in the PLBs after incubation in different concentrations of sucrose for different periods of time determined by HPLC. The highest sucrose concentration was 7 mg/g of PLBs for the PLBs treated with 0.75 M sucrose for 3 days as compared to the control which had only 1 mg/g sucrose. After sucrose preculture, the PLBs were subjected to desiccation using one of two methods. Desiccation using silica gel was more efficient in reducing PLBs moisture content. After 6 h of desiccation, PLBs desiccated using laminar air flow had 43.5% moisture content while for those desiccated using silica gel had 32% moisture content. PLBs desiccated to different moisture contents were plunged into LN. After storage in LN the encapsulated PLBs were re-warmed. Two weeks after re-warming PLBs viability was determined by TTC reduction and re-growth assessed. Encapsulated PLBs precultured with 0.75 M sucrose for 3 days followed by desiccated using silica gel for 5 h resulting in a moisture content of 39% lead to the highest post re-warming viability in terms of TTC reduction (46.6% of control PLBs) and 30% re-growth.