不同天然居群小檗种子萌发障碍因子研究

以西天山野果林霍城居群、新源居群和特克斯居群黑果小檗以及特克斯居群红果小檗种子为试验材料,研究了4组野生小檗种子生物学特性、吸水特性以及去种皮、低温层积和不同浓度GA₃处理对小檗种子休眠与萌发特性的影响,结果表明：（1）4组野生小檗种子的吸水率均表现出逐渐增加的趋势,黑果小檗种子与红果小檗种子吸水率差异不明显,其种皮透水透气性相似;（2）霍城居群、新源居群和特克斯居群黑果小檗种子的种皮对萌发存在强烈的抑制作用;而新源居群红果小檗种子的种皮抑制作用不明显;（3）4℃低温层积处理对4组野生小檗种子萌发影响很大,随层积时间的增加小檗种子发芽率均逐渐提高,3组黑果小檗种子层积90 d时,休眠基本被解除;红果小檗种子层积30 d时,休眠基本被解除;（4）浓度为200 mg·L^-1的GA₃溶液处理可显著提高4组小檗种子发芽率,浓度过高或过低均会对小檗种子萌发起到抑制作用。层积60 d时霍城居群、新源居群和特克斯居群黑果小檗去种皮种子用200 mg·L^-1的GA₃溶液处理2 h后,发芽率分别为85.00%、83.33%和86.67%;红果小檗层积15 d时去种皮种子用浓度为200 mg·L^-1的GA₃溶液处理后,发芽率可达86.67%。研究结果将为今后野生小檗的种质资源保护,利用及可持续发展提供技术支持和科学依据。     

短瓣金莲花种子无菌萌发与幼苗快繁技术初探

以野生短瓣金莲花种子为材料,探讨不同处理方式对种子发芽的影响,并比较不同培养基类型及激素组合对短瓣金莲花根茎部位形成的愈伤组织、不定芽增殖及生长的影响,建立无菌播种及组培快繁体系,为短瓣金莲花快繁和规模化生产提供技术支撑。结果表明：（1）用600 mg·L^-1 GA₃溶液浸泡48 h后并用0.1% HgCl₂溶液灭菌20 min的种子发芽效果最好;（2）适宜于增殖的培养基配方为MS+0.5 mg·L^-1 6-BA+0.1 mg·L^-1 NAA;（3）适宜于生根的培养基配方为不添加激素的MS。     

胡萝卜人工种子包衣材料的筛选

以胡萝卜SK4-316种子苗下胚轴为外植体诱导培养的体细胞胚,进行人工种子的制作。结果表明,海藻酸钠包埋体系制作的人工种子萌发率较稳定,PEG体系的萌发率高于前者,但萌发后成苗率较低。以MS液体培养基作包埋培养基,4%海藻酸钠、2% CaCl₂·2H₂O、0.1 mg/L GA₃、0.2%活性炭组成的海藻酸钠复合包衣剂制作的人工种子,在1/16MS培养基、24℃下培养萌发率最高,可达97.02%。     

檀香种胚离体培养快速育苗研究

利用添加植物生长调节剂的培养基对檀香种胚进行离体培养,研究种胚萌发和成苗的影响因素以及种苗快速繁育技术。结果显示:生长调节剂GA₃和种胚成熟度都是影响檀香种胚萌发和成苗的重要因素,檀香成熟胚在含14 μmol/L GA₃的MS培养基上7~10 d即可萌发,45 d后成苗率达82.5%,檀香种胚离体培养是有效的快速育苗方法。     

寄主植物根浸提液对肉苁蓉种子萌发及吸器形成的影响

利用滤纸培养皿法研究藜科植物梭梭和甜菜根水浸提和乙醇浸提两种浸提液对肉苁蓉种子萌发和吸器形成的影响。结果表明: 水和乙醇浸提的根浸提液对肉苁蓉种子萌发和吸器形成无明显作用;两种根醇浸提液添加10 mg·kg^-1赤霉素(GA₃)处理的肉苁蓉种子萌发率均提高了10倍以上,但与只加赤霉素处理组(GA₃对照)无显著差异。在根浸提液中添加1 mg·kg^-1氟啶酮(FL)处理的肉苁蓉种子萌发率与FL对照组无显著差异,其中甜菜根水浸提液处理组肉苁蓉种子萌发率最高,达39.4%。与根浸提液中添加赤霉素处理仅能使萌发率提高不同,寄主根浸提液中添加FL后,萌发肉苁蓉种子芽管上均有吸器形成,梭梭根醇浸提处理的吸器形成率最高,达16.2%。梭梭根醇浸提液中同时添加GA₃和FL,肉苁蓉种子萌发率可提高到52.3%,但吸器形成率与浸提液中添加FL处理无差异;FL对照仅有6.7%的萌发肉苁蓉种子形成吸器,显著低于梭梭根醇浸提液添加FL处理组。不同处理的肉苁蓉种子吸器形成位置和形态有差异,添加寄主根浸提液处理的吸器大多出现在芽管顶端,多个乳头状凸起成爪状;未添加寄主根浸提液的FL对照组吸器出现位置大多在芽管底部或顶端出现分叉。研究证明,乙醇浸提和水浸提两种方式都能从寄主根中提取出促进肉苁蓉种子吸器形成的物质但对促进种子萌发作用不明显;GA₃和FL可显著提高肉苁蓉种子的萌发率,但萌发肉苁蓉种子吸器的形成受寄主根浸提液中某些物质的影响。     

温水浸种对杜仲种子萌发过程中保护酶活性及内源激素含量变化的影响

探讨杜仲（Eucommia ulmoides Oliver）种子萌发过程中保护酶活性及内源激素含量的变化，为其种子破眠及萌发不同时期采取有效处理措施，从而提高出苗率提供参考。利用室温（CK）、40℃、50℃、60℃和70℃不同水温，探讨温水浸种对杜仲种子萌发的影响。种子萌发进程中，利用常规生理生化方法测定种子内超氧化物歧化酶（SOD）、过氧化物酶（POD）活性和丙二醛（MDA）含量，以及酶联免疫法测定赤霉素（GA₃）、吲哚乙酸（IAA）、脱落酸（ABA）和玉米素核苷（ZR）含量。结果表明：①温水浸种处理能提高了杜仲种子的发芽率及发芽势，其中以50℃温水处理最为显著。②与室温浸种处理相比，50℃温水浸种处理提高了种子内SOD及POD活性，减轻了质膜氧化损伤程度，致使MDA含量的持续下降，从而促使种子朝向有利于萌发的方向发展。此外，50℃温水浸种处理在提高种子内促进生长内源GA₃、IAA及ZR含量的同时，并降低了抑制生长的内源ABA含量，致使GA₃/ABA、IAA/ABA和ZR/ABA值均明显高于室内浸种处理，最终促进了杜仲种子的萌发。     

Biological activities of gibberellins and their glucosides in Pharbitis nil

Growth-promoting effects of gibberellins and their glucosides isolated from immature seeds of Japanese morning glory (Pharbitis nil) were compared in six bioassay systems. GA₃ glucoside exhibited much less activity than GA₃ in the dwarf rice (under aseptic conditions), dwarf maize (d₁, d₂ and d₅), cucumber and dwarf pea assays. GA₈, GA₂₆, GA₂₇ and GA₂₉ showed low activities in all the bioassay systems, while their glucosides were even less active. Thus gibberellin glucosides do not appear to be active in growth regulation.     

Desiccation decreases abscisic acid content in hybrid larch (Larix X leptoeuropaea) somatic embryos

Previous studies indicated that the high endogenous abscisic acid (ABA) content of hybrid larch ( Larix X leptoeuropaea ) somatic embryos was correlated with low germination frequency. However, when dried, the germination rate of the somatic embryos improved. Therefore, our present objective was to study the effect of desiccation on the levels of ABA and its glucose ester metabolite. Cotyledonary somatic embryos were subjected to drying treatments at 4⁰C under relative humidities of 98 and 59% for one week and the levels of both ABA and abscisic acid glucose ester (ABAGE) were followed by enzyme-linked immunoassay (ELISA). During desiccation at 98% relative humidity (RH) both ABA and ABAGE levels decreased in an irregular fashion. Regardless of RH, transient increases in ABA were observed that were paralleled by marked decreases in ABAGE. It is concluded that the desiccation of somatic embryos which leads to a decrease in ABA content, could explain the enhanced germination capacity of such embryos.     

粗茎秦艽种子粗提物与破眠方法研究

为了探究影响粗茎秦艽（Gentiana crassicaulis Duthie ex Burk.）种子休眠的因素,破除休眠,寻找其种子快速萌发的方法,以干燥的粗茎秦艽种子为材料,测定种子吸水率及粗提物的活性,使用不同浓度的赤霉素（GA₃）、高锰酸钾（KMnO₄）、聚乙二醇（PEG6000）和过氧化氢（H₂O₂）溶液进行浸种处理,比较不同处理条件对粗茎秦艽种子萌发的影响。结果显示,粗茎秦艽种皮对种子吸水没有明显阻碍作用;不同浓度的种子粗提物对白菜、小麦的萌发和生长均表现出抑制作用;不同浓度的粗提物对粗茎秦艽种子自身的萌发也表现出一定的抑制作用,当粗提物浓度达到0.1 g/mL时,抑制作用最显著（P < 0.05）;高锰酸钾处理可提高粗茎秦艽种子的萌发率,浓度为1.5%时效果较显著（P < 0.05）,而过氧化氢处理对粗茎秦艽种子的萌发效果不如前者,此外,用500 mg/L的赤霉素浸种和300 mg/L的聚乙二醇预处理也可显著打破粗茎秦艽种子休眠（P < 0.01）。研究结果表明粗茎秦艽种子的内源抑制物是影响其休眠的因素之一;种皮的机械阻碍也在一定程度上影响了种子萌发;粗茎秦艽种子具有综合性休眠特性。高锰酸钾预处理、赤霉素浸种和聚乙二醇引发均可打破种子休眠、缩短种子出芽时间,提高种子的发芽能力。     

Increase in permeability to oxygen and in oxygen uptake of soybean nodules under limiting phosphorus nutrition

In vitro cultures and endogenous abscisic acid (ABA) analyses with gas chromatography-selected ion monitoring-mass spectrometry (GC-SIM-MS) were used to study the effects of AgNO₃ and polyethylene glycol (PEG) on white spruce ( Picea glauca ) somatic embryo maturation and endogenous ABA contents, Normally, in the absence of ABA, white spruce somatic embryos cannot mature. However, AgNO₃ and PEG stimulated somatic embryo maturation by increasing the number of cotyledonary embryos. A combination of 100 μ M AgNO₃ and 40 g l⁻¹ PEG was the treatment that was most effective in enhancing cotyledonary embryo formation without exogenous ABA. Either AgNO₃ or PEG was able to increase endogenous ABA levels of the embryogenic culture but a combination of AgNO₃ and PEG was the most effective treatment. Stimulation of cotyledonary embryo formation by AgNO₃ occurred only at low ABA levels, while PEG promoted embryo formation at all exogenous ABA concentrations. Germination tests indicated that AgNO₃ had no negative effects on embryo germination and conversion while the PEG-treated embryos failed to germinate.     

解除休眠处理对凤丹种子萌发和幼苗生长的影响

凤丹种子具有很深的休眠特性。本实验以AgNO₃、GA₃、KT、TDZ和4℃低温处理凤丹种子,探讨解除其上胚轴休眠的途径。结果表明,AgNO₃、GA₃和低温(4℃)处理能加快凤丹种子萌发并提高发芽率,且不同程度地改善幼苗的鲜重、株高、根系等生长指标。     

枇杷叶片胚性愈伤组织诱导与内源激素含量的关系

为揭示胚性愈伤组织发生过程中内源激素的变化规律,本研究以枇杷叶片为实验材料,通过诱导胚性愈伤组织获得体细胞胚,采用高效液相色谱法测定枇杷叶片及愈伤组织的赤霉素（GA₃）、生长素（IAA）、脱落酸（ABA）和玉米素（ZT）4种内源激素的含量,探讨胚性愈伤组织发育过程中4种内源激素的动态变化。结果显示,不同成熟度叶片内IAA/ZT比值对胚性愈伤组织的发生有正效应,而GA₃/IAA的比值具有负效应。胚性愈伤组织的发生需要较低含量的GA₃及高含量的IAA和ABA,IAA/ZT比值高有利于胚性愈伤组织形成,培养后期及时添加一定量的外源激素有利于胚性保持。本研究可为离体培养时选择外植体、添加外源激素及控制继代时间提供理论指导,并为快速获得枇杷胚性材料、开展基因转化研究奠定基础。     

Solid substrate cultivation of Gibberella fujikuroi on an inert support

Growth of Gibberella fujikuroi on Amberlite, an inert support, and gibberellic acid (GA₃) production was studied in glass columns under different conditions of temperature and water activity (a_w). Maximum biomass concentration and GA₃ production were respectively 40 (mg/g inert support) and 0.73 (mg/g inert support). While high specific growth rates were obtained, low initial nitrogen resulted in low biomass concentrations. Maximum GA₃ (31°C, a_w=0.985) was not produced by the maximum concentration of biomass (25°C, a_w=0.992). Peaks in the rate curves of either outlet gas, CO₂ or O₂, occurred on exhaustion of urea indicating, for future works, just when to feed the culture additional nitrogen.     

寄生植物锁阳种子萌发方法及愈伤组织、初生吸器诱导研究

专性寄生植物锁阳以干燥肉质茎入药,为常用中药材。在锁阳的生活史中,需要经历种子萌发、芽管状器管伸长、初生吸器形成、次生吸器形成等过程,其中种子萌发和初生吸器形成是锁阳完成生活史的最基本条件。目前,触发锁阳种子萌发、初生吸器形成的植物生长调节物质的阈值和种类并不清楚,导致人工调控锁阳生活史困难及栽培收益不高。本论文运用组织培养方法,研究赤霉素、生长素和细胞分类素等多种激素交互作用对锁阳种子萌发和吸器形成的影响。研究结果显示：（1）多种激素的共同作用促进锁阳球形原胚的发育。（2）B₅培养基添加1.0 mg·L^-1 2,4-D,0.5 mg·L^-1 KT,1.0 mg·L^-1 GA₃可以高效诱导锁阳种子愈伤组织形成,愈伤诱导率达13.7％±3.1%。（3）B₅培养基添加0.5 mg·L^-1 2,4-D,0.25 mg·L^-1 KT可以高效诱导锁阳愈伤组织分化初生吸器,一些初生吸器继续分化成芽管状气管,延伸3~4 cm后,顶端膨大,形成新的初生吸器。本论文研究结果可为进一步研究锁阳种子萌发、初生吸器形成的内源激素变化规律及发生机制研究奠定基础。     

理化因子对龙血树柴胡种子萌发的影响

本研究以龙血树柴胡（Bupleurum dracaenoides Huan C.Wang,Z.R.He&H.Sun）的种子为材料,采用不同温度、不同浓度赤霉素、不同化学试剂以及紫外线照射和微波辐射等方式处理后,统计其发芽率、发芽势、发芽指数和发芽时间等指标。结果表明：龙血树柴胡种子萌发最适温度为20℃;100 mg/L的GA₃、0.5％的KMnO₄和3％的H₂O₂处理均能促进其种子萌发,但不能使种子萌发和出苗时间提前;25 s微波辐射处理可大幅提高种子发芽率;紫外线照射处理种子0.5 h,萌发效果最佳,但随照射时间的延长对种子萌发则有不同程度的抑制。     

3,4-二羟基苯乙酮胁迫对天山云杉种子萌发过程中内源植物激素含量变化的影响

3,4-二羟基苯乙酮(DHAP)是天山云杉(Picea schrenkiana ssp. tianschanica)叶和凋落物中存在的主要自毒物质, 是导致天山云杉林天然更新障碍的原因之一。为了解释自毒物质发生作用的生理机制, 该文设计多个浓度梯度的DHAP溶液处理天山云杉种子, 以发芽率和发芽势为种子萌发参数, 运用反相超高效液相色谱(UPLC)分析技术, 检测了种子萌发过程中内源植物激素玉米素(ZT)、赤霉素(GA₃)、吲哚乙酸(IAA)和脱落酸(ABA)含量水平的变化。研究结果表明: DHAP处理对天山云杉种子萌发影响具有浓度效应, 表现为5.0 mmol·L^-1 > 0.1 mmol·L^-1 > 1.0 mmol·L^-1 >对照, 即5.0 mmol·L^-1 DHAP处理组对种子萌发的抑制作用最强、0.1 mmol·L^-1 DHAP处理组次之、1.0 mmol·L^-1 DHAP处理组最弱; DHAP处理组的种子内源ZT、GA₃浓度水平降低, ABA含量升高, GA₃浓度峰值出现时间延迟, IAA浓度在高浓度(5.0 mmol·L^-1 DHAP)处理组短时间内(3-6天)过量积累, 1.0和5.0 mmol·L^-1 DHAP处理组的种子内源ABA浓度峰值出现时间延迟; DHAP处理种子萌发1-6天时, ZT/(GA₃+IAA)比值降低, IAA/ZT、ABA/ZT比值增大; ABA/(ZT + GA₃ + IAA)比值在0.1 mmol·L^-1 DHAP处理组增大, 在5.0 mmol·L^-1 DHAP处理组降低。DHAP处理引发种子内源激素含量水平及激素含量间比值变化, 可能是抑制、延迟天山云杉种子萌发的主要原因。     

Induction of tolerance to fast desiccation in black spruce (Picea mariana) somatic embryos: relationship between partial water loss,sugars, and dehydrins

Events associated with the induction of tolerance to fast desiccation in black spruce ( Picea mariana ) somatic embryos were investigated. An experimental approach using an initial period of partial water loss was developed to induce either no, partial, or complete tolerance to fast desiccation. Tolerance to subsequent fast desiccation was not promoted by decreasing embryo water content from 1.5 to 1.1 g H₂O g⁻¹ DW (g g⁻¹) throughout the first 24 h of slow desiccation. However, tolerance increased from 10 to 95% germination during the second 24-h period of slow desiccation after partial water loss from 1 to 0.55 g g⁻¹. Emphasis was also placed on the relationship between observed tolerance, and sugar and dehydrin contents. Compared to controls, sucrose content in embryos doubled after 24 h of slow desiccation and more than tripled after 48 h. Conversely, starch content was decreased by one half after 24 h and by three quarters after 48 h. Sucrose abundance and raffinose occurrence after 48 h of slow desiccation were congruent with complete tolerance to fast desiccation. The period of slow desiccation between 24 and 48 h also increased the content of a 24-kDa dehydrin and the appearance of a 42-kDa dehydrin. The relationship between partial water loss, sugars and dehydrins is discussed with respect to tolerance to fast desiccation in black spruce somatic embryos.     

Enhancing carrot somatic embryos survival during slow dehydration, by encapsulation and control of dehydration

In order to obtain dry artificial seeds, carrot somatic embryos were encapsulated and dehydrated. Encapsulation in some hydrogels delayed the dehydration and preserved the water content of carrot somatic embryos. In particular, a matrix made of alginate with gellan gum was found to be the most efficient in maintaining a high water activity (a_w) around somatic embryos. By delaying dehydration, and also rehydration, encapsulation seemed to protect somatic embryos against desiccation and imbibition damages, giving better germination and emergence of cotyledons. Matrices made of alginate mixed with kaolin or gellan gum were particularly adapted to protect the embryos during the dehydration. Apart from the matrix composition, the control of dehydration speed enhanced the survival and regeneration of encapsulated-dehydrated somatic embryos. Using a slow dehydration protocol (95-15% RH—relative humidity into the chamber—in 11.5 days), it was possible to exert different dehydration speeds. Slowing the dehydration between 70 and 45% RH stabilized the water activity (a_w) of the encapsulation matrix, and enhanced the survival and regeneration frequencies of encapsulated-dehydrated embryos. In the absence of any maturing pretreatment, alginate-gellan gum encapsulated carrot somatic embryos, dehydrated to 15% RH, and rehydrated in moistured air (90% RH), germinated up to 72.9%. Therefore, encapsulation in alginate-gellan gum, combined with a slow dehydration, leads to enhance the somatic embryos' desiccation tolerance.     

不同栽培措施对降香黄檀内源激素含量的影响

为了了解不同栽培措施对降香黄檀叶片内源激素的影响,本研究通过铺设修枝、移植、钾肥、乙烯4个试验,分析各处理叶片激素含量特征。结果表明：修枝实验中,重度修枝处理IAA、GA₃、ZR、ABA含量均最大,分别为69.849、6.619、8.805、77.998 ng·g^-1。移植各处理中,去冠移处理IAA、ZR含量最高分别为73.195和9.472 ng·g^-1,而其ABA含量最低为52.001 ng·g^-1,GA₃含量最高的为断根处理8.418 ng·g^-1。IAA、ZR、GA₃含量K1钾肥配比最高分别为75.188、8.383、6.127 ng·g^-1;ABA最大含量71.082 ng·g^-1为CK处理。乙烯试验中,CK处理的IAA、GA₃含量最高分别为47.762、4.967 ng·g^-1;E2.5%处理的ABA含量96.94 ng·g^-1最高。E0.1%处理的ZR含量最高为9.378 ng·g^-1。修枝对GA₃/ABA和ZR/GA₃影响较大,其最小值分别比最大值降低了29.7%、19.0%。移植对IAA/ABA、GA₃/ABA影响较大,最小值比最大值依次降低了52.47%和51.47%。树干注射乙烯和钾肥对IAA/ABA、GA₃/ABA影响较大,乙烯试验最小值分别比最大值降低了55.4%、49.2%,钾肥试验最小值分别比最大值降低了48.8%和37.1%。