Pollen and seed desiccation tolerance in relation to degree of developmental arrest, dispersal, and survival

In most species, arrest of growth and a decrease in water content occur in seeds and pollen before they are dispersed. However, in a few cases, pollen and seeds may continue to develop (germinate). Examples are cleistogamy and vivipary. In all other cases, seeds and pollen are dispersed with a variable water content (2-70%), and consequently they respond differently to environmental relative humidity that affects dispersal and maintenance of viability in time. Seeds with low moisture content shed by the parent plant after maturation drying can generally desiccate further to moisture contents in the range of 1-5% without damage and have been termed 'orthodox'. Pollen that can withstand dehydration also was recently termed orthodox. Seeds and pollen that do not undergo maturation drying and are shed at relatively high moisture contents (30-70%) are termed 'recalcitrant'. Since recalcitrant seeds and pollen are highly susceptible to desiccation damage, they cannot be stored under conditions suitable for orthodox seeds and pollen. Hence, there are four types of plants with regard to tolerance of pollen and seeds to desiccation. Orthodoxy allows for dispersal over greater distances, longer survival, and greater resistance to low relative humidity. The advantage of recalcitrance is fast germination. Orthodoxy and recalcitrance are often related to environment rather than to systematics. It has been postulated that certain types of genes are involved during presentation and dispersal of pollen and seeds, since molecules (sucrose, polyalcohols, late embryogenic abundant proteins, antioxidants, etc.) that protect different cell compartments during biologically programmed drying have been detected in both.     

The effects of some storage conditions on viability of Lecanicillium lecanii conidia to whitefly (Homoptera: Trialeurodes vaporariorum)

A study on the survival of Lecanicillium lecanii conidia in storage at room temperature was carried out. Firstly, drying methods of conidia powder were compared. Vacuum-freeze drying (VFD) was more suitable for drying conidia as compared to vacuum drying (VD) at room temperature. Vacuum-freeze drying for 24-h resulted in a water content of 5.4%, and a viability, determined as germination of conidia in 2% glucose solution after16 h, was 90.3% and the infection in greenhouse whitefly, Trialeurodes vaporariorum was about 94.7% at a dose of 1×10⁸ conidia/mL. Secondly, the factors influencing viability of conidia stored at room temperature were evaluated in the laboratory. Temperature was the most critical factor influencing conidial storage stability, among the tested factors affecting survival of conidia stored at room temperature for 6 months. Both conidial germination and infection of hosts decreased with storage temperature increasing from 15 to 35°C, and at 35°C the survival of stored conidia for 6 months was near zero. The moisture content of the conidial powder was another major factor influencing viability of stored conidia at room temperature. Conidial powder dried to about 5% moisture content showed higher viability than non-dried conidial powder. For the carriers, clay and charcoal were more suitable for storage of L. lecanii conidia at room temperature. At a room temperature of 25°C, L. lecanii conidia which were dried to 5% water content and mixed with clay or charcoal could retain about 50% survival after 6 months' storage.     

不同干燥方式、存储温度对羊栖菜中岩藻黄素稳定性的影响

为探究不同干燥方式、不同存储温度和存储时间对羊栖菜中岩藻黄素稳定性的影响,本研究考察了三种不同干燥方式,包括自然阴干、低温烘干和冷冻干燥,对羊栖菜中岩藻黄素稳定性的影响。结果表明采用冷冻干燥所得羊栖菜,含水量低,岩藻黄素含量高,达到699.2μg/g,岩藻黄素保留率为94.7%,远高于自然阴干及低温烘干。因此,冷冻干燥是最佳干燥方法。考察了四个不同温度(-20、4、25、30℃)存储对冻干羊栖菜中岩藻黄素稳定性的影响,结果表明随着存储温度的升高,岩藻黄素的降解率明显增高,因此,-20℃为最佳储存温度。     

Effect of drying on conidial viability of Penicillium frequentans, a biological control agent against peach brown rot disease caused by Moniliniaspp

The effects of drying methods (freeze-, spray-, and fluid bed-drying) on viability of Penicillium frequentans conidia were compared. Viability, estimated by germination of fluid bed- and freeze-dried conidia, was similar to that of fresh conidia. Skimmed milk alone, or in combination with other protectants, was added to conidia before freeze-drying. After the freeze-drying process, all protectants used, except glycerol improved conidial viability. Freeze-dried P. frequentans conidia did not maintain viability after 30 days of storage at room temperature, while conidia dried by fluid bed-drying showed 28% viability following 180 days after drying. This work also demonstrated a relationship between conidial viability after 1 year of storage at room temperature, moisture content after fluid bed-drying and initial weight of sample. Conidial moisture contents must be reduced to 5-15% for optimal storage at room temperature. P. frequentans conidia dried by fluid bed-drying were as effective as fresh conidia in controlling brown rot of peaches.     

Improved Equations for the Prediction of Seed Longevity

Equations for predicting seed longevity in storage have beenimproved so that they now take into account variations withina species in initial seed quality—which is affected bygenotype and pre-storage environment—and so that theyare more accurate over a wider range of storage environmentsThese improvements have been incorporated into a seed viabilitynomograph for barley (Hordeum distichum L.) which may be usedto predict percentage viabihty of any seed lot after any timein any storage environment within the range –20 to 90°C and 5–25 per cent moisture content. Applicationsof the improved equations to seed drying and to long-term seedstorage for genetic conservation are discussed. Hordeum distichum L., barley, seed viability, seed longevity prediction, seed storage, seed drying, storage temperature, seed moisture content, genetic resources conservation     

桃花粉低温和超低温保存方法比较研究

桃(Prunus persica(L.)Batsch)是我国重要的无性繁殖作物种质资源,目前主要保存于3个国家无性繁殖作物种质圃。随着以茎尖、花粉、休眠芽为保存载体的超低温保存技术的发展,超低温保存已成为无性繁殖作物重要备份保存方式。本研究以15份桃种质花粉为研究对象,开展含水量、回湿处理和保存温度(4℃低温保存和液氮超低温保存)对保存后花粉离体萌发率的影响研究。研究结果:明确了桃种质花粉超低温保存的含水量;揭示了回湿处理对部分桃种质花粉超低温保存产生显著影响;超低温保存后花粉离体萌发率最高可达83%;4℃低温保存和超低温保存比较研究结果表明,超低温保存4年后14份桃种质花粉离体萌发率仍可保持30%以上,11份桃种质花粉离体萌发率与保存前花粉离体萌发率相比无显著变化甚至显著提高,而4℃低温保存的花粉离体萌发率降至0。该研究为国家种质库建立花粉规模化超低温保存提供技术支撑。     

Viability of Lettuce Seeds: I. SURVIVAL IN HERMETIC STORAGE

Seeds which show orthodox storage characteristics conform toa common pattern of survival over a wide range of storage conditionswhich can be described by a single equation. Two aspects ofthis are that in a constant environment the life-spans of individualseeds in a population are normally distributed, and that thereis negative linear relationship between log moisture contentand log life-span. However, when orthodox seeds are fully hydratedthey survive much longer than would be predicted by extrapolatingfrom lower moisture contents; but the moisture content at whichthe change occurs has not previously been investigated. In this paper the viability of lettuce seeds was examined aftervarious periods of hermetic storage at different moisture contents,temperatures and initial partial pressures of oxygen. At moisturecontents below 15% the pattern of survival is typical of otherorthodox seeds but above this value the responses change infour ways: instead of being deleterious, oxygen becomes beneficialto survival; instead of life-spans being normally distributed,they become skewed; the relative effect of temperature on decreasinglongevity is slightly diminished; and the decrease in eurvivalperiod with increase in moisture content begins to become lessmarked, so that ultimately, above 20% to 30% moisture content,there is no further decrease in longevity. These results, which indicate substantial physiological changesat about 15% moisture content, are discussed in relation tothe hypothesis, postulated by Villiers, that repair and turnovermechanisms are absent from dry seeds but are activated on hydration. Key words: Lettuce, Lactuca saliva L., Seed viability, Seed storage     

Long-term storage of Pennisetum glaucum (L.) R. Br. pollen

Summary Pearl millet [Pennisetum glaucum (L.) R. Br.] pollen has been successfully stored for 2,615 and 2,911 days at -18° and -73 °C, respectively, and continues to be viable. Viability of pollen stored at -73 °C appears to be little affected either by pollen storage moisture contents below 7.2% or by storage in glass vial or zip-lock plastic bag containers. Pollen moisture content appears to be more critical for maintaining viability at -18°C than at -73°C. Glass vials appear to be more desirable for longer term (>3 years) storage at -18°C.     

不同贮藏温度下大别山五针松花粉活力的变化

大别山五针松( Pinus dabeshanensis C. Y. Cheng et Y. W. Law)为松科( Pinaceae)松属( Pinus Linn.)植物,自然种群数量极少,目前仅发现在安徽省岳西县大王沟海拔900~1300 m阴坡和半阴坡有相对集中的分布,种群规模计200余株,且多为成年个体,林下幼苗极少,自然更新困难,为中国特有珍稀树种之一[1]。大别山五针松常在每年3月份至4月份形成花芽,5月中下旬花粉成熟并散发,花粉具气囊,此时雌球花张开接受传粉,球果翌年9月成熟[2]。据作者近年的调查,大别山五针松种子败育率较高,且种子质量较低,这也是该种类濒危的重要原因之一。     

Fluidized-bed drying and storage stability of Cryptococcus flavescens OH 182.9, a biocontrol agent of Fusarium head blight

A method to produce dried granules of Cryptococcus flavescens (formerly Cryptococcus nodaensis) OH 182.9 was developed and the granules evaluated for storage stability. Small spherical granules were produced and dried using a fluidized-bed dryer. A drying and survival curve was produced for the process of fluidized-bed drying at 30°C. The granules were dried to different moisture contents (4, 7, 9 and 12%) and evaluated for storage stability at 4°C for up to a year. These different moisture contents granules had the following respective water activities (0.22, 0.38, 0.47 and 0.57 a _w). The results show the storage stability varied significantly across this moisture content range. The 9% moisture content sample had the best short-term stability (up to 4 months), while 4% moisture content had the best long-term survival (1 year). A desorption isotherm of C. flavescens was determined and modeled. The results of the storage stability and drying studies are interpreted in context of the desorption isotherm.     

Storage behavior of Typha latifolia pollen at low water contents: Interpretation on the basis of water activity and glass concepts

Germplasm must be stored under optimal conditions to maximize longevity and efficiently maintain genetic resources. In order to identify optimal storage conditions, we investigated the effects of temperature (−5 to 45°C) and water content (<0.17 g H₂O g⁻¹ dry weight) on longevity of Typha latifolia L. pollen. Longevity was highest at water contents corresponding to storage relative humidity (RH) of 11‐15% which corresponded to the shoulder of water sorption isotherms. Also coinciding with this shoulder were abrupt changes in heat capacity of water present in the pollen. Consistent with changes in isotherms with temperature and the concept of critical RH for storage, optimum water contents increased with decreasing temperature. An attempt was made to explain the aging behavior according to the glass concept. The water content‐temperature combinations of optimal storage were found to be below the glass transition curve, indicating that optimum storage conditions are achieved when intracellular glasses are present. We also found a change in activation energy of aging in Arrhenius plots around T_g, demonstrating a change in aging kinetics when the glassy state is lost. We concluL that T_g curves cannot be used solely to predict precise conditions of optimum storage, but might be useful for predictions of storage longevity above optimum water contents. The data imply that too much drying reduces longevity and should be avoided, particularly when cryogenic storage is considered.     

Viability of rape (Brassica napus L.) seeds following selection on the basis of newly-emerged radicles then subsequent drying and storage

Germinating rape seeds selected on the basis of newly-emerged radicles (1 ± 0.5 mm) were dried to an equilibrium moisture content (c. 11%) in air at 20°C and 80% relative humidity without loss of viability. Storage life of these low-moisture-content germinating (LMCG) seeds at 15°C was limited to 7 days before viability was significantly reduced. However, viability of LMCG seeds was maintained for 84 days in storage at -20°C. Longer periods in store reduced viability, but 96% of seeds still remained viable after 336 days at - 20°C. Increasing periods of storage at -20°C reduced the subsequent seed longevity at 15°C, indicating a reduction in vigour during storage. Storage under reduced pressure or in a nitrogen atmosphere had little significant effect on seed longevity. Reduction of moisture content below 11% using vacuum drying at a range of temperatures reduced seed vigour.     

Microflora of Barley Kernels

Numbers and kinds of microflora were determined in 160 samples of barley grown in different regions of the United States; microflora were more abundant in the grains grown in the central states than in those grown in the western states. During steeping and germination in micromalting equipment, the number of colonies of filamentous fungi increased from two to five times, colonies of yeasts from five to ten times, and bacteria from 50 to more than 100 times the numbers present in the grain before malting. Kiln drying according to a commercial schedule reduced the number of all types of microflora below the number present before kilning, but all were present in larger numbers in the kilned malt than in the original grain. In barley stored at room temperature and at a moisture content of 15 to 18%, members of the Aspergillus glaucus group increased with increasing time and increasing moisture content, and germination percentage of the seeds decreased. Stored free of storage fungi at room temperature, barley with a moisture content just over 15% retained a high germination percentage for 5 months, but at a moisture content of 16% the germination decreased to zero.     

The Effect of Temperature and Moisture Content on the Longevity of Seed of Ulmus carpinifolia and Terminalia brassii

Seeds of the Smooth-leafed Elm (Uimus carpinifolia) and of thetropical forest tree Terb (Terminalia brassii) were stored hermeticallyand sampled at intervals for periods of up to two years. Bothspecies possess ‘orthodox’ seed (increasing longevityis observed as either moisture content or temperature are reduced)within the temperature ranges from — 13 to 52°C (Elm)and from —4 to 42°C (Terb) and within the moisturecontent ranges from 3 to 19 per cent (Elm) and from 5 to 14per cent (Terb) on a fresh weight basis. Elm seed stored at—75°C showed the expected relationship between longevityand moisture content, but did not differ significantly in longevityfrom seed kept at — 13°C when moisture contents wereheld constant. Probit analysis of the relationship between germinationpercentage and time was performed for each storage environment,yielding a slope from which the standard deviation of the distributionof seed deaths over time () was calculated. Standard deviationvalues were used in turn to determine the values of constantsin a viability equation which had previously been applied toseed of barley, chickpea, cowpea and soybean. The equation,which gave a good fit to the results obtained, can be used topredict viability for seed in storage over a wide range of environmentalconditions. Some limitations to the applicability of the viability equationwere defined. At 22 per cent and higher moisture contents Elmseed survived longer than predicted. Furthermore, all Elm andTerb seed was killed quickly on placing in —75°C at22 and 20 per cent moisture content respectively, but high viabilitywas retained for several days at 19 and 17 per cent respectively.Practical implications of the results are discussed. Uimus carpinifolia Gleditsch, Smooth-leafed Elm, Terminalia brassii Exell, Terb, seed longevity, seed storage, moisture content, temperature     

Microflora of Barley Kernels

Numbers and kinds of microflora were determined in 160 samples of barley grown in different regions of the United States; microflora were more abundant in the grains grown in the central states than in those grown in the western states. During steeping and germination in micromalting equipment, the number of colonies of filamentous fungi increased from two to five times, colonies of yeasts from five to ten times, and bacteria from 50 to more than 100 times the numbers present in the grain before malting. Kiln drying according to a commercial schedule reduced the number of all types of microflora below the number present before kilning, but all were present in larger numbers in the kilned malt than in the original grain. In barley stored at room temperature and at a moisture content of 15 to 18%, members of the Aspergillus glaucus group increased with increasing time and increasing moisture content, and germination percentage of the seeds decreased. Stored free of storage fungi at room temperature, barley with a moisture content just over 15% retained a high germination percentage for 5 months, but at a moisture content of 16% the germination decreased to zero.     

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。     

蓝花丹开花生物学观测及花粉储存研究

对四川引种栽培的蓝花丹（Plumbago auriculata Lam.）花粉与柱头形态、花部特征及开花物候进行了系统观测,并对其花粉的采集时间与储存温度进行了初步研究。结果显示：（1）两种类型花粉和柱头形态均具有较大差异,短花柱型（S型）花粉的极轴长P值和赤道轴长E值均显著大于长花柱型（L型）花粉,L型柱头则明显长于S型,且两种花粉的纹饰和柱头的瘤状突起物形态也明显不同;（2）蓝花丹具有较高的开花同步性指数（为0.89）,但相对开花强度不高,属于中等强度;（3）蓝花丹两种花型在花蕾半开放时采收花粉,其活力最高,L型花粉活力可达85.24%±4.22%,S型可达87.74%±2.95%;（4）L型与S型花粉分别于25℃干燥0.5 h和1 h后再低温储存,其活力保持更好;（5）干燥后的花粉在-86℃条件下保存效果最好,储存30 d后L型花粉活力高达66.51%±0.85%、S型达69.07%±1.57%。本研究表明蓝花丹二型植株在生殖资源分配上存在明显差异,这种差异一方面导致花粉大小和形态及其所需干燥时间的不同,另一方面导致二型花柱头表现出明显不同的特征,这些差异性的结构是否参与了蓝花丹自交不亲和反应的识别过程,还有待进一步研究。此外,较低的开花强度会造成虫媒传粉困难,这可能是蓝花丹自然结实率极低的重要因素之一。     

A Model of the Effect of Temperature and Moisture on Pollen Longevity in Air-dry Storage Environments

Data on the survival of pollen ofTypha latifoliaL. stored forup to 261 d over seven different saturated salt solutions (providing0.5 to 66% relative humidity) and six different constant temperatures(from -5 to +45 °C) were analysed to quantify the effectof air-dry storage environment on pollen longevity. Pollen survivalcurves conformed much more closely to negative cumulative normaldistributions than to negative exponential relations. Estimatesofp₅₀(storage period required to reduce pollen viability to50%), provided by negative cumulative normal distributions,were available from 37 different storage environments in whichpollen viability was reduced below 50%. Once observations at0.5% and 5.5% relative humidity were excluded from analysis,there was a negative logarithmic relation between these estimatesof longevity and pollen moisture content (%, wet basis) anda curvilinear semi-logarithmic relation between longevity andtemperature. When the negative logarithmic relation betweenlongevity and moisture content was replaced by a negative semi-logarithmicrelation between longevity and the relative humidity of thestorage environment the resultant model was less satisfactory,principally because pollen longevity over saturated solutionsof calcium nitrate (43–62% relative humidity) and sodiumnitrite (60–66% relative humidity) were consistently greaterand smaller, respectively, than fitted values. Notwithstandingthese errors, comparison between the fitted relations and observationsat the two lowest relative humidities provided estimates ofthe lower-relative-humidity limits to these relations. Theseprovisional estimates varied with storage temperature beinglowest at 25 °C (<5.5% relative humidity). However, therewas no linear trend to that variation (P>0.25): the meanestimate was 11.9 (s.e.=1.4)%. The considerable similaritiesamong models of pollen longevity in air-dry storage, and theirestimated lower limits, and those developed previously for orthodoxseeds and spores are discussed.Copyright 1999 Annals of BotanyCompany. Typha latifoliaL., pollen, storage, survival, longevity, relative humidity, moisture content, temperature.     

Differential desiccation sensitivity of corn and Pennisetum pollen linked to their sucrose contents

Abstract. Two pollen species from the Gramineae family were investigated as to their differential sensitivity to desiccation. Germination in vivo ceased to occur after previous drying to 7–8% moisture content in Zea mays and 3% in Pennisetum typhoides. The reduced vitality coincided with extensive membrane damage, as evidenced by the considerable leakage of fluorescein and K⁺ from the prehumidified grains into liquid media. Further experiments were undertaken to investigate the basis for this interspecific difference: (1) phospholipid analyses revealed little difference in composition and content both before and after drying; (2) free fatty acid contents increased in both species upon drying but levels were generally low; and (3) sucrose was the sole soluble carbohydrate found in both species. Fresh maize pollen contained 5% sucrose, as compared to 14% in Pennisetum pollen. During slow drying (8h) this level went up to 12% and 17%, respectively. Drying corn pollen in the cold (2°C), or at high rate, limited sucrose levels and affected the resistance to drying. The authors conclude that survival of dehydration is correlated with the presence of sucrose. Finally, they tested the ability of sucrose to preserve dry liposomes prepared from phospholipids purified from the two species. When liposomes were dried in the presence of sucrose, fusion and leakage could largely be prevented at mass ratio's of sugar to lipid of > 4. Trehalose was also effective but myo-inositol was not. No species differences were observed. The authors suggest that the presence of sucrose is a key factor in preserving membranes in dry pollen.     

The effects of moisture and temperature on the ageing kinetics of pollen: interpretation based on cytoplasmic mobility

This study shows that characterization of the molecular mobility in the cytoplasm of pollen provides a new understanding of the effects of moisture and temperature on ageing rates. Using EPR spectroscopy, we determined the rotational motion of the polar spin probe, 3-carboxy-proxyl, in the cytoplasm of Typha latifolia pollen, under different temperature and moisture content conditions. Increasing the temperature resulted in faster rotational motion, analogous to faster ageing rates. With decreasing moisture content, rotational motion first decreased until a minimum was reached, after which rotational motion slightly increased again. The moisture content at which this minimal rotational motion was observed increased with decreasing temperature, comparable to the pattern of ageing rate. A significant linear relationship was found between ageing rates and rotational motion in the cytoplasm, suggesting that these parameters are causally linked. Upon melting of the intracellular glass, a twofold increase in activation energy of rotational motion and ageing rate was observed. In contrast, melting of the sucrose glass resulted in an increase in rotational motion of five orders of magnitude. The difference in rotational motion upon melting glasses of pollen or sucrose suggests that other molecules beside sugars play a role in intracellular glass formation in pollen.