种子超干贮藏技术应用面临的问题和研究方向

种子超干贮藏技术对种质资源保存具有很大的应用前景,但鉴于目前的研究进展,其应用还存在诸多技术上的问题。本文对种子超干贮藏技术在种子库中应用所面临的挑战以及需要开展的研究内容进行了如下几个方面的讨论：种子贮藏安全含水量下限的多样性决定了超干技术应用的复杂性;超干贮藏含水量安全下限的确定尚未有可靠的方法;超干贮藏的种子吸胀前需要恰当的预处理以避免吸胀损伤;超干处理前外源抗氧剂预处理可能改善超干贮藏效果;无氧条件可能对超干贮藏具有增效作用;糖类物质以及两性物质对种子在超干处理及超干贮藏过程中的保护作用。     

超干种子的膜功能与糖组分的研究

采用两相法分离种子质膜,研究超干处理对种子质膜ATP酶活力及膜流动性的影响.结果表明,白菜(Brassica pekinensis(Lour.)Rupr.)、榆树(Ulmus pumila L.)种子经超干处理后,在人工老化和自然老化条件下,与在-20℃条件下贮藏的种子比较,超干贮藏种子质膜ATP酶活力和微粘度无显著变化,过于贮藏种子的效应虽有所下降,但仍比高含水量室温贮藏的对照种子为好.此结果与超干种子具较高活力水平完全一致,说明超干贮藏种子保持了质膜的生理功能,因此提高了种子的耐藏性.高效液相色谱分析结果显示超干种子中还原糖/非还原糖的比值低于高含水量种子,积累的蔗糖、水苏糖含量与超干种子的耐干力有关.玉米(Zea mays L.)种子中不含水苏糖,这可能是玉米种子较其他种子耐干力下降的原因之一.     

洋葱种子含水量与贮藏温度对其寿命的影响

不同含水量（MC7.1％～1.2％）的洋葱种子贮藏在35℃、室温、15℃和5℃条件下1～3年,适度超干处理能延长种子的贮藏寿命;种子的贮藏寿命与种子含水量和贮藏温度密切相关。种子贮藏的最适含水量随温度的改变而发生相应的变化,35℃时MC为3.4％;室温时为3.4％～4.5％;15℃时为4.5%～5.1％。MC≤2.2％不利于延长种子寿命。在室温自然条件下贮藏1～3年,适度超干种子（MC3.4％）内MDA和H2O2含量、O－*2产生速率和LOX活性明显地低于未超干种子（MC7.1％）和高度超干种子（MC1.2％）,而抗氧化酶AsA-POD、CAT和SOD的活性显著地高于未超干种子（MC7.1％）和高度超干种子（MC1.2％）。据此认为对脂质过氧化的抑制作用是适度超干种子耐贮藏的生理原因之一。     

超干种子的膜功能与糖组分的研究

采用两相法分离种子质膜,研究超干处理对种子质膜ATP酶活力及膜流动性的影响。结果表明。白菜（Brassica pekinensis(Lour.)Rupr)。榆树（Ulmus pumilaL）种子经超干处理后,在人工老化和自然老化条件下,与在-20℃条件下贮藏的种子比较,超干贮藏种子质膜ATP酶活力和微粘度无显著变化,过干贮藏种子的效应虽有所下降,但仍比高含水量室温贮藏的对照种子为好。此结果与超干种子具较高活力水平完全一致。说明超干贮藏种子保持了质膜的生理功能。因此提高了种子的耐藏性。高效液相色谱分析结果显示超干种子中还原糖/非还原糖的比值低于高含水量种子,积累的蔗糖,水苏糖含量与超干种子的耐干力有关。玉米（Zea maysL.)种子中不含水苏糖,这可能是玉米种子较其他种子耐干力下降的原因之一。     

洋葱种子含水量与贮藏温度对其寿命的影响

不同含水量（MC 7．1％－1．2）的洋葱种子贮藏在35℃、室温、15℃和5℃条件下1－3年,适度超于处理能延长种子的贮藏寿命;种子的贮藏寿命与种子含水量和贮藏温度密切相关。种子贮藏的最适含水量随温度的改变而发生相应的变化,35℃时MC为3．4％;室温时为3．4％－3．5％;15℃时为4．5％－5．1％。MC≤2．2％不利于延长种子寿命。在室温自然条件下贮藏1－3年,适度超干种子（MC3．4％）内MDA和H2O2含量、O2^-产生速率和LOX活性明显地低于未超干种子（MC7．1％）和高度超干种子（MC1．2％）,而抗氧化酶AsA-POD 、CAT和SOD的活性显著地高于未超干种子（MC7．1％）和高度超干种子（MC1．2％）。据此认为对脂质过氧化的抑制作用是适度超干种子耐贮藏的生理原因之一。     

超干贮藏榆树种子萌发过程中ATP和可溶性糖含量的变化

经过超干(种子含水量3.73%)贮藏(普通室温下2个月)的榆树种子萌发过程中ATP与可溶性糖含量和ATP酶、淀粉酶、抗氧化酶的活性都高于对照(种子含水量9.34%,普通室温下贮臧),说明适度的超干贮藏有利于保持种子活力.     

超干贮藏榆树种子萌发过程中ATP和可溶性糖含量的变化

经过超干（种子含水量3．73％）贮藏（普通室温下2个月）的榆树种子萌发过程中ATP与可溶性糖含量和ATP酶,淀粉酶,抗氧化酶的活性都高于对照（种子含水量9．34％,普通室温下贮藏）,说明适度的超干贮藏有利于保持种子活力。     

超干贮藏杜仲种子的研究

杜仲种子含水量降至３．５％以下,其耐藏力增强。经５０℃１０天人工加速老化后,表现出较强的抗老化能力。室温贮藏２年后,与５℃、－２０℃条件贮藏种子相比,发芽率、根长及活力指数等均无显著性差异。超干种子比对照种子（含水量７．５％,室温贮藏）电导率降低４５％;脂质过氧化产物丙二醛（ＭＤＡ）的积累减少;对脱氢酶活性的保持能力比常规种子增强。超干种子幼苗的吸氧量高于对照种子。经过氧化物酶同工酶谱分析,超干种     

超干处理提高榆树种子的耐藏性（简报）

短命的榆树种子,自然贮藏1至2个月失去萌发力,而超干处理后其含水量降至2％以下,活力变化不大。室温下贮藏3个月的种子活力仍很高。超干贮藏与低温贮藏的效果一致。     

贮藏温度和超干处理对沙芥属种子萌发及解剖结构的影响

以沙芥种子(含水量4.5%)和斧形沙芥种子(含水量4.3%)为材料,采用硅胶干燥法将其含水量分别降至3.1%、2.2%、1.3%,在50℃、35℃和20℃条件下密闭贮藏1年,研究贮藏温度和超干处理对种子萌发指标、解剖结构的影响,为种子老化及贮藏研究提供理论参考。结果表明:(1)随着贮藏温度升高和种子含水量降低,沙芥和斧形沙芥种子的发芽率、胚根长、胚根干质量均在下降。(2)解剖观察发现,贮藏温度升高和种子含水量降低,首先加剧种子胚内部维管束组织活性的降低,其次由里及外对薄壁细胞活性和结构产生破坏,同时胚淀粉粒活性和数量下降。(3)沙芥和斧形沙芥种子分别在含水量4.5%和4.3%(CK)、并于20℃～35℃条件下贮藏后,种子的萌发和结构活性保持最好,而且斧形沙芥种子的保持能力强于沙芥。     

Starch Mobilization in Ultradried Seed of Maize (Zea mays L.) During Germination

The effects of ultradry storage on the starch mobilization in maize (Zea mays L.) seed after aging were investigated. The results indicated that there were no significant differences in the content of ATP,starch, and soluble sugar, as well as the activity of amylase, between ultradried seeds and seeds stored at -20 ℃ during germination. These results were consistent with the higher level of vigor of the ultradried seed. Sieve tube introduction of a fluorescence dye (carboxyl fluoresceindiacetate) and laser confocal microscopy were used to study the development of plasmodesmata in the ultradried seeds. The results indicated that plasmodesmata developed well in ultradried seeds. Fluorescence analysis also showed that the fluorescence intensity in the radicle of ultradried seeds was stronger than that in seeds with a higher moisture content. This suggests that ultradry treatment has no adverse effects on the seeds. After seed imbibition, cell orgaelles could be resumed. It is concluded that ultradry seed storage is beneficial for maintaining seed vigor and that starchy mobilization proceeds regularly during germination.     

Starch Mobilization in Ultradried Seed of Maize （Zea mays L.） During Germination

The effects of ultradry storage on the starch mobilization in maize (Zea mays L.) seed after aging were investigated. The results indicated that there were no significant differences in the content of ATP, starch, and soluble sugar, as well as the activity of amylase, between ultradried seeds and seeds stored at -20℃ during germination. These results were consistent with the higher level of vigor of the ultradried seed. Sieve tube introduction of a fluorescence dye (carboxyl fluoresceindiacetate) and laser confocal microscopy were used to study the development of plasmodesmata in the ultradried seeds. The results indicated that plasmodesmata developed well in ultradried seeds. Fluorescence analysis also showed that the fluorescence intensity in the radicle of ultradried seeds was stronger than that in seeds with a higher moisture content. This suggests that ultradry treatment has no adverse effects on the seeds. After seed imbibition, cell orgaelles could be resumed. It is concluded that ultradry seed storage is beneficial for maintaining seed vigor and that starchy mobilization proceeds regularly during germination.     

超干种子耐贮藏性的细胞学及生理生化基础

种子超干贮藏是近年来种子生物学研究和植物种质资源保存领域的研究热点。本文综述了种子超干贮藏研究的进展 ,包括 :(1)超干种子的细胞和亚细胞结构变化 ;(2 )超干处理对种子水分状态的影响以及与劣变反应速率的关系 ;(3)超干种子中的自由基伤害和有毒物质的积累 ;(4)超干种子中的内源抗氧剂 ;(5 )糖类物质以及两性物质对极度脱水种子细胞的保护。此外 ,还对此领域的研究进行了展望     

含水量对种子贮藏寿命的影响

建立以收集种子为主体的基因库乃是当今保护植物种质资源最为普遍且可靠易行的方式,在世界库存约 61 0 0 0 0 0份种质资源中,近 90 %是以种子形式保存于约 1 30 0个基因库中。低温贮藏仍是目前基因库中种子种质保存的主要方法。种子含水量和贮藏温度是影响种子在贮藏期间生活力和活力保持的关键因素。传统的经验认为控制温度比控制水分来得安全有效,因而趋向于向低温或超低温的贮藏方向发展。国际植物遗传资源研究所（ＩＰＧＲＩ）曾推荐 5%～ 6%的含水量和 - 1 8～- 2 0℃低温作为各国长期保存种子的理想条件。目前,世界各国都把更多的…     

超干对种子活力及水分热力学特性的影响

以耐干的菜心种子和不耐干的豇豆种子为材料,研究了超干对其活力和水分吸附等温线及热力学参数（△S、△H、△G）的影响。超干使红豇豆种子活力下降,SOD、POD活性降低,膜结构受损,这些变化即发生在水分降低的干燥过程中又发生在眼干状态下的贮藏过程中,菜心种子的活力和SOD,POD的活性受超干的影响小,超干对细胞膜的轻微损伤可在萌发吸胀时得到修复,超干种子水分吸附等温线上区段1和区段2的含水量明显下降;超干种子除－△S和－△H在区段1略有所升高外,－△S、－△H在区段和2和区段3和－△G在各区段均有程度不同的下降;这些改变尤以豇豆种子较显著,说明豇豆种子的水分束缚能力低,较易失去水分吸附等温线上的强吸附水,继而引起大分子结构的改变,降低种子活力;而菜心种子则相反,能在超干状态下较好地保持酶的活性和细胞膜的结构,故适宜超干贮藏。     

Theoretical Basis of Protocols for Seed Storage III. Optimum Moisture Contents for Pea Seeds Stored at Different Temperatures

In previous work, we demonstrated that there was an optimummoisture level for seed storage at a given temperature (Vertucciand Roos, 1990), and suggested, using thermodynamic considerations,that the optimum moisture content increased as the storage temperaturedecreased (Vertucci and Roos, 1993b). In this paper, we presentdata from a two year study of aging rates in pea (Pisum sativum)seeds supporting the hypothesis that the optimum moisture contentfor storage varies with temperature. Seed viability and vigourwere monitored during storage under dark or lighted conditionsat relative humidities between 1 and 90%, and temperatures between-5 and 65°C. The optimum moisture content varied from 0·015g H₂O g^-1 d.wt at 65°C to 0·101 g H₂O g^-1 d.wt at15°C under dark conditions and from 0·057 at 35°Cto 0·092 g H₂O g^-1 d.wt at -5°C under lighted conditions.Our results suggest that optimum moisture contents cannot beconsidered independently of temperature. This conclusion hasimportant implications for 'ultra-dry' and cryopreservationtechnologies.Copyright 1994, 1999 Academic Press Seed storage, seed aging, seed longevity, water content, temperature, glass, desiccation damage, ultradry, Pisum sativum L., pea, cryopreservation     

我国作物种质资源保存研究与展望

我国于１９８４年１９８６年在北京建成两座国家种质库,９０年代初又在青海建立一座复份库,形成了国家库中,长期配套和异地结合的种质资源保存体系。截止１９９３年底,国家库保存种质资源达２７万余份,到１９９５年保存总数将达到３０余万份。与种子入库时,在国家库的管理,种子质量检测,种子干燥以及低温保存技术研究等方面取得一定进展,今后将继续扩大种质贮存、加强贮存种子生活力跟踪监测,无破坏性检测技术,贮存生理和     

Relationship of chlorophyll, seed moisture and ABA levels in the maturing Brassica napus seed and effect of a mild freezing stress

Chlorophyll (Chl) retention by mature seed of canola as the result of an early frost or other environmental factors (the "green seed problem") causes serious economic losses. The relationship of seed degreening to rate of moisture loss by seed and silique and the role of ABA in this process were investigated as a function of developmental age. During the normal predesiccation stage (28–45 days after pollination), seed of Brassica napus (cv. Westar) loses Chl rapidly but seed moisture slowly. After a mild freezing stress, there is a rapid loss of moisture from silique walls, followed by accelerated loss of seed moisture. Chl degradation ceases at 35–45% seed moisture. ABA levels in silique walls of frozen plants (determined by enzyme‐linked immunosorbant assay) increased after freezing, apparently in response to moisture loss. In contrast, ABA levels in the seed increased dramatically 1 day after freezing, then decreased to control levels. The influence of the rate of seed moisture loss on Chl degradation was investigated by fast and slow drying of isolated seed under controlled humidity conditions. Seed dried rapidly at 22% RH retained most of its Chl, whereas seed dried slowly at 86% RH lost Chl as fast or faster than seed on control (unfrozen) plants. In all treatments, Chl loss stopped at about 40% seed moisture.     

Loss of Viability in Lettuce Seeds and the Accumulation of Chromosome Damage under Different Storage Conditions

Loss of seed viability in lettuce (Lactuca sativa L.) duringstorage is associated with an increase in the frequency of cellsin the surviving seeds showing chromosome damage during firstmitoses. The relation is linear when probit of the frequencyof aberrant cells is plotted as a function of probit percentagenormal germination. The slope of the relation, however, variesaccording to moisture content so that the proportion of aberrantcells for any given loss of germination increases with decreasein moisture content over the range 13.0–5.5 per cent.At 3.3 per cent moisture content, however, the proportion ofaberrations was no greater than at 5.5 per cent moisture content;and at 18.1 per cent moisture content the proportion was noless than at 13.0 per cent moisture content. Despite these differences,the increase in chromosomal aberrations per unit time for agiven temperature was always less the lower the moisture content.Diplontic selection markedly reduced the frequency of chromosomalaberrations and eliminated the differences in these frequenciesbetween the different storage treatments. But even after fiveweeks' growth, root tips from aged seed still contained abouttwice as many aberrant cells as compared with similar root tipsderived from the original seed stock. Studies on the frequencyof recessive mutations indicated that excessive amounts of heritablemutations were not present in the progenies of aged seed, evenwhen stored at moisture contents as low as 5.5 per cent. Allthis and other evidence reinforces the view that orthodox seedsfor genetic conservation should be stored at not more than about5 per cent moisture content, and that even lower moisture contentsare worth considering. The results also emphasise the need formaintaining a high regeneration standard, i.e. the percentageto which seed viability is allowed to fall during storage beforethe seed stock is regenerated. Lactuca sativa, lettuce, seed storage, seed viability, chromosomal aberrations, phenotypic mutations     

The effect of time of harvest on the viability and pre-emergence mortality in soil of pea (Pisutn sativum L.) seeds

A series of harvests from field plots of peas showed that the younger seeds were at the time of harvest the lower was their percentage viability and the greater their percentage mortality in soil. Young seed also showed a high degree of solute leaching into steep water and had a high moisture content at harvest compared with older seeds. Heavy rainfall just prior to harvesting reduced the percentage viability of the dried seed and increased the moisture content of the seed at harvest as well as the leaching of solutes from the dried seed into steep water. Comparisons of moisture content at harvest and seed condition after drying suggested that cotyledons and embryo axes were only able to withstand enhanced drying after removal from the plant if the seed moisture content had already begun to decline whilst the seed was still on the plant.