秸秆还田对旱作马铃薯块茎淀粉合成关键酶活性及基因表达的影响

为探讨秸秆还田下旱作马铃薯块茎形成过程中淀粉合成关键酶活性及基因表达特性,以马铃薯栽培品种"青薯9号"为材料,以露地栽培为对照,设置秸秆还田处理,研究了马铃薯块茎形成过程中淀粉合成关键酶活性、基因表达、淀粉糊化及累积指标。结果表明:秸秆还田显著提高了旱作马铃薯SSS酶活性,降低了AGPP、GBSS酶活性,而对SBE酶活性没有显著影响;显著提高了SSⅡ、SSⅢ基因表达量,降低了AGPase、GBSSⅠ、SBEⅠ、SBEⅡ基因表达量;显著增加了淀粉崩解值,减少了淀粉各阶段粘度、回生值,而对淀粉糊化温度没有显著影响;显著增加了直链淀粉含量及直/支链淀粉比,减少了总淀粉含量;GBSS酶活性与AGPase、SBEⅠ基因表达量呈显著正相关,与直链淀粉含量、直/支链淀粉比呈显著负相关;SBE酶活性与SSⅡ基因表达量、峰值粘度、低谷粘度、最终粘度、总淀粉含量呈显著正相关,与崩解值、糊化温度呈显著负相关;AGPase基因表达量与直链淀粉含量呈显著负相关;GBSSⅠ基因表达量与最终粘度、回生值呈显著正相关,与糊化温度呈显著负相关;淀粉糊化与累积无显著相关性。     

秸秆还田对旱作马铃薯块茎淀粉合成关键酶活性及基因表达的影响

为探讨秸秆还田下旱作马铃薯块茎形成过程中淀粉合成关键酶活性及基因表达特性,以马铃薯栽培品种"青薯9号"为材料,以露地栽培为对照,设置秸秆还田处理,研究了马铃薯块茎形成过程中淀粉合成关键酶活性、基因表达、淀粉糊化及累积指标。结果表明:秸秆还田显著提高了旱作马铃薯SSS酶活性,降低了AGPP、GBSS酶活性,而对SBE酶活性没有显著影响;显著提高了SSⅡ、SSⅢ基因表达量,降低了AGPase、GBSSⅠ、SBEⅠ、SBEⅡ基因表达量;显著增加了淀粉崩解值,减少了淀粉各阶段粘度、回生值,而对淀粉糊化温度没有显著影响;显著增加了直链淀粉含量及直/支链淀粉比,减少了总淀粉含量; GBSS酶活性与AGPase、SBEⅠ基因表达量呈显著正相关,与直链淀粉含量、直/支链淀粉比呈显著负相关; SBE酶活性与SSⅡ基因表达量、峰值粘度、低谷粘度、最终粘度、总淀粉含量呈显著正相关,与崩解值、糊化温度呈显著负相关; AGPase基因表达量与直链淀粉含量呈显著负相关;GBSSⅠ基因表达量与最终粘度、回生值呈显著正相关,与糊化温度呈显著负相关;淀粉糊化与累积无显著相关性。     

兰州百合组培鳞茎发育研究

该研究以兰州百合商品种球鳞片为外植体材料,通过组织培养诱导丛生芽萌发及高频增殖,再以丛生芽为材料诱导其发育形成小鳞茎,调节培养基对小鳞茎进行膨大发育培养,最终形成促进兰州百合组培鳞茎膨大发育的“三步”组培培养技术路线;对发育过程中形成的丛生芽、小鳞茎及膨大鳞茎进行淀粉含量测定与生长特征参数统计,分析各步培养对鳞茎形成发育过程中淀粉含量与形态变化的影响。结果表明：所建立的“三步”培养方案培育出的组培鳞茎直径、重量与鳞片数分别为1.66 cm、2.48 g和26.33片,有效地促进了鳞茎的膨大,并能诱导鳞茎主茎杆的形成发育;在培养进程中其淀粉含量呈现逐步增加的趋势,这表明与鳞茎膨大发育正相关,同时鳞茎大小、重量及鳞片数三者也表现为正相关性;当鳞茎所含鳞片数在26片以上时,其生长点易发育形成主茎杆。该文研究了兰州百合组培鳞茎的形成与膨大发育技术,所研发的“三步”培养组成的鳞茎膨大发育组培技术有效地促进了鳞茎的膨大发育,而膨大发育的鳞茎能有效地缩短田间生长周期,从时间上提高百合生产量,同时为实现兰州百合膨大的鳞茎种球规模化生产提供技术支撑。     

兰州百合组培鳞茎发育研究（英文）

该研究以兰州百合商品种球鳞片为外植体材料,通过组织培养诱导丛生芽萌发及高频增殖,再以丛生芽为材料诱导其发育形成小鳞茎,调节培养基对小鳞茎进行膨大发育培养,最终形成促进兰州百合组培鳞茎膨大发育的"三步"组培培养技术路线;对发育过程中形成的丛生芽、小鳞茎及膨大鳞茎进行淀粉含量测定与生长特征参数统计,分析各步培养对鳞茎形成发育过程中淀粉含量与形态变化的影响。结果表明:所建立的"三步"培养方案培育出的组培鳞茎直径、重量与鳞片数分别为1.66 cm、2.48 g和26.33片,有效地促进了鳞茎的膨大,并能诱导鳞茎主茎杆的形成发育;在培养进程中其淀粉含量呈现逐步增加的趋势,这表明与鳞茎膨大发育正相关,同时鳞茎大小、重量及鳞片数三者也表现为正相关性;当鳞茎所含鳞片数在26片以上时,其生长点易发育形成主茎杆。该文研究了兰州百合组培鳞茎的形成与膨大发育技术,所研发的"三步"培养组成的鳞茎膨大发育组培技术有效地促进了鳞茎的膨大发育,而膨大发育的鳞茎能有效地缩短田间生长周期,从时间上提高百合生产量,同时为实现兰州百合膨大的鳞茎种球规模化生产提供技术支撑。     

百合鳞茎发育过程中碳水化合物含量及淀粉酶活性变化

以兰州百合和亚洲系"精粹"百合为试材,探讨了鳞茎发育过程中不同部位淀粉、可溶性糖含量和淀粉酶活性的变化。结果表明,母鳞茎作为百合萌发阶段的代谢源,其外部鳞片是代谢更为活跃的部位。淀粉和可溶性糖含量同时增加是百合新鳞茎开始膨大的标志。蔗糖是百合鳞茎中可溶性糖的主要形态,还原糖的变化体现了碳水化合物的供应及转化。淀粉酶在百合鳞茎发育过程中对调节和平衡碳水化合物的形态起重要作用。     

AGPase及其基因表达研究

ADP-葡萄糖焦磷酸化酶(AGPase)是淀粉生物合成的限速酶,本文讨论了到目前为止对不同植物AGPase基因及其表达的研究结果。其中包括AG-Pase大小亚基编码基因表达的分布及调控。其结论为:完整AGPase两个大亚基和两个小亚基对合成正常含量的淀粉缺一不可;有多个等位基因编码植物AGPase大小亚基,它们表现发育阶段、组织器官、细胞质体内外及诱导条件的分化表达;尽管各亚基表达于含淀粉的组织,但它们在储藏组织内的表达量远高于其他组织;对调控不敏感的外源AGPase基因导入淀粉植物将会成为提高淀粉含量的重要途径。提出了进一步研究中存在的问题及可能的解决途径。     

葛根淀粉合成关键酶活性动态及其与块根产量和淀粉积累的相关性研究

为探讨葛根发育过程中淀粉合成关键酶活性与块根产量和淀粉积累的关系,以初步揭示其内在的生理机制。该研究以‘桂葛1号’粉葛和‘桂葛8号’野葛为材料,采取生理测定法对农艺性状、直链和支链淀粉的含量、淀粉合成关键酶活性等进行测定,并对葛根发育过程中淀粉合成关键酶活性、农艺性状和淀粉含量动态变化的关系进行相关性分析。结果表明：(1)块根发育过程中,两品种葛根腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)、可溶性淀粉合成酶(SSS)、束缚性结合淀粉合成酶(GBSS)和淀粉分支酶(SBE)的活性呈现先增大后降低的单峰曲线变化,与直链淀粉、支链淀粉和总淀粉含量由缓慢增长到快速增长后趋于稳定的变化趋势基本一致,即在块根形成期至膨大期逐渐增长,至膨大后期达到最大,之后迅速下降,至成熟期缓慢下降,并维持在较高水平。(2)‘桂葛8号’的淀粉含量和产量显著高于‘桂葛1号’,其酶活性也均显著高于‘桂葛1号’。(3)葛块根的根长、根粗、单株重、干物质含量、产量表现为“缓慢-快速-稳定”的变化趋势,淀粉含量表现出类似变化。(4)相关性分析结果显示,4个淀粉合成关键酶活性与块根直链淀粉、支链淀粉及总淀粉含量、根长均呈显著或...     

复合调控Wx与SBE3基因对水稻籽粒直链淀粉含量的影响

颗粒淀粉合成酶（GBSS）和淀粉分支酶3（SBE3）是淀粉合成过程中的两个关键酶,这两个酶主要由耽和SBE3两个基因分别控制,它们的表达量直接影响直链淀粉和支链淀粉的含量比例。为了探讨水稻淀粉关键酶基因耽过量与SBE3干涉复合表达对直链淀粉含量的影响,构建了Wx过量表达与SBE3干涉结合的多基因表达载体,并通过农杆菌介导的方法将其导入日本晴水稻中。经过PCR检测分析获得了65株转基因阳性植株,半定量RT—PCR检测表明转基因株系中Wx基因表达量明显增加,而SBE3基因表达量显著减少。转基因株系籽粒透明度明显降低,直链淀粉含量比野生型的平均高45％,但是千粒重变化不大,与野生型相当。遗传分析表明这些转基因株系多数可稳定遗传。     

转反义Wx基因水稻颖果中与淀粉合成和积累相关酶活性的变化

将反义Wx基因转入水稻,导致Wx蛋白不同程度减少,颖果中的直链淀粉含量不同程度下降,总淀粉含量显著降低,直链淀粉与总淀粉的比值极显著降低。在水稻颖果发育过程中,ADPG-PPase、GBSS、SSS和SBE的活性在灌浆前期迅速升高,达最大值后很快下降,在灌浆中后期下降趋缓。Wx蛋白减少后的转基因水稻颖果中的GBSS活性明显下降,下降幅度与直链淀粉含量相一致,而且活性高峰期比其亲本有所提前。转基因水稻颖果中ADPG-PPase和SSS的活性在颖果发育的前中期,SBE则在中后期高于相应的亲本。     

细叶百合LpPEX7基因克隆及盐胁迫下的表达特性分析

从细叶百合的鳞茎中克隆出过氧化物酶体生物合成蛋白基因（LpPEX7）,该基因ORF全长957 bp,编码318个氨基酸。LpPEX7蛋白序列包含6个WD40保守结构域,通过同源蛋白序列比对和进化树分析,发现LpPEX7与其他植物的PEX7蛋白具有较高的同源性。LpPEX7基因在细叶百合种子,叶片和鳞茎中的表达量比较高,在根和花中表达量比较低,在H₂O₂,NaCl,NaHCO₃不同逆境处理条件下,LpPEX7基因的表达量都发生了改变。在盐碱和氧化胁迫处理下,LpPEX7过表达拟南芥株系种子的萌发要早于野生型种子的萌发,这些研究结果表明LpPEX7基因与盐碱、氧化逆境有一定的应答关系,为细叶百合的耐盐碱性分子机理研究提供一个非常重要的候选基因。     

Stimulating effect of spermine on bulblet formation in bulb-scale segments of Lilium longiflorum

When bulb-scale segments of Lilium longiflorum were cultured on a medium containing auxin and cytokinin, the proportion of the expiants with newly-formed bulblets was significantly increased by the application of different polyamines. The most effective polyamine was spermine, where more than 90% of segments formed an average of 5 bulblets as compared to controls where less than 50% explants formed an average of 1.5 bulblets. Application of arginine one of the precursors putrescine biosynthesis, slightly promoted bulblet formation. The putrescine-stimulated bulblet formation was strongly inhibited by simultaneous addition of an inhibitor of the spermidine synthase, cyclohexylamine. The spermidine-promoted bulblet formation, however, could not be suppressed by this inhibitor. The promotive effect of spermidine on bulblet formation was reversed by an inhibitor of the spermine synthase, N-(3-aminopropyl)cyclohexylamine, but application of this inhibitor with spermine did not show any apparent effect on the bulblet formation. Endogenous level of spermine increased in common during bulblet formation that were stimulated by exogenous polyamines. Thus, spermine seemed to be the main stimulating chemical on bulblet formation in lily bulb-scale segments.Abbreviations APCHA N-(3-aminopropyl)cyclohexylamine - Arg arginine - BA benzyladenine - CHA cyclohexylamine - MS Murashige and Skoog's - NAA naphthaleneacetic acid - Orn ornithine - Put putrescine - Spd spermidine - Spm spermine     

Differential Effects of Paclobutrazol on the Bulblet Growth of Oriental Lily Cultured In Vitro: Growth Behavior,Carbohydrate Metabolism,and Antioxidant Capacity

Bulblet growth is crucial for global lily production, and the applications of plant growth regulators have proven effective but with poor understanding of mechanisms. The aims of this study were to identify the effects of paclobutrazol (PBZ) on tube lilies. Low PBZ concentrations (5 × 10⁻⁴ mM, LPBZ) stimulated bulblet biomass, whereas higher doses suppressed the growth of the leaves and roots. Soluble carbohydrate and starch contents increased significantly with increased PBZ dose. The activities of adenosine 5′-diphosphate glucose pyrophosphorylase (AGPase), granule-bound starch synthase (GBSS), and soluble starch synthase (SSS) increased dramatically in response to PBZ treatments at later growth stages (60 days after transplanting) when carbon starvation occurred. In contrast, GBSS activity was enhanced throughout the whole growth period, indicating that the starch increase was attributed mainly to amylose synthesis. Carbohydrates were utilized more efficiently following LPBZ, with a relative bulblet weight of approximately 77.07%, which potentially ensured the source-sink balance. Alternatively, more carbohydrates were stored in response to high PBZ concentration. Intriguingly, PBZ usage significantly increased the activities of ascorbate peroxidase, catalase and glutathione reductase at the early stages, implicating a possible role in the elimination of ROS to maintain homeostasis. Application of LPBZ resulted in the largest bulblet, which weighed 396 mg (2.5 times that of the control) and measured 10.70 mm in diameter. The detailed characterizations of the bulblet swelling mechanism using PBZ in vitro offer suggestions of future PBZ usage in other bulbous crops.

     

<Emphasis Type="Italic">In vitro</Emphasis> regeneration and bulblet growth from lily bulbscale explants as affected by retardants,sucrose and irradiance

Bulbscales of oriental lily hybrid Star Gazer were used as the explants. Bulblets were formed on the basal portion of the excised bulbscales on MS medium supplemented with growth retardants, different sucrose concentrations and exposed to continuous light or dark. ^{Alar, Cycocel} and Paclobutrazol in concentration 1 mg dm⁻³ produced higher number of bulblets as compared to the control. The number of bulblets, however, decreased with the increase in concentration of the growth retardants. The number of bulblets was higher at 90 than at 60 g dm⁻³ sucrose and when the bulbscales were exposed to continuous light than to darkness. The growth retardants, higher sucrose concentration and continuous dark stimulated fresh mass of bulblets. The number of bulblets having roots and leaves decreased in medium with Alar, Cycocel and Paclobutrazol as compared to the control. A few bulblets produced roots and leaves in medium with 90 g dm⁻³ sucrose and none of the regenerated bulblets produced leaves under continuous dark.     

In vitro propagation from axenic explants of <Emphasis Type="Italic">Lilium oxypetalum</Emphasis> (D. Don) Baker,an endemic bulbous plant of high altitude Himalaya

In vitro propagation protocol for Lilium oxypetalum, a high altitude Himalayan endemic lily, has been developed. Effect of explant types (i.e., callus and in vitro bulblet scales) and sucrose concentration [0–6.0% (w/v)] on in vitro bulblet regeneration of L. oxypetalum was tested in previously optimized Murashige and Skoog basal medium supplemented with 2.0 μM 6-benzyladenine and 0.1 μM α-naphthaleneacetic acid. Callus explants produced significantly (P < 0.01) higher number of bulblets per explant than bulblet scale explants. Of the different concentrations of sucrose tested, 4.5% (w/v) sucrose showed significantly (P < 0.01) higher percentage regeneration (i.e., 70.8 ± 4.2 and 79.2 ± 4.2% regeneration on callus and bulblet scale explants, respectively), and produced higher number of bulblets per explant (i.e., 9.0 ± 0.4 and 5.4 ± 0.5 bulblets on callus and bulblet scale explants, respectively). Regenerated bulblets developed 2–3 leaves when subcultured for 4 weeks and were subsequently transferred ex vitro with a survival rate of 66.7% after 6 weeks. Leaves of the survived plantlets became dry after growing ex vitro for 10 weeks, amongst which 86.4% re-sprouted after remaining dormant for 5–6 weeks and produced 1.5 bulblets per explant. Findings of the present study hold promise for efficiently multiplying the target species in view of its potential economic and conservation significance.     

Lilium candidum bulblet and meristem development

Lilium candidum L., commonly known as the Madonna lily, is a wild Lilium species with medicinal properties and excellent potential as an ornamental crop, but one that has been scarcely investigated. The aim of this research was to study (1) tissue culture propagation of L. candidum bulblets, (2) early bulblet development, and (3) the effect of temperature and bulblet weight on bulblet and plant growth and meristem development. An investigation of the effect of explant type and temperature on in vitro bulblet propagation showed that scales were the most efficient explants for in vitro propagation and that exposing the regenerating bulblets to 15°C for 4 wk increased bulblet weight but reduced the number of bulblets produced. For bulblets planted in soil after 12 wk of exposure to 15°C or 25°C, the fastest growth was observed in the bulblets that had been exposed to 15°C and that had a larger initial size. Histological examination showed that young in vitro-grown bulblets had a rudimentary meristem comprising few cells with no layer organization. After 12 wk of growth, all bulblets showed a layered meristem, regardless of bulblet size or exposure to 15°C. However, an increased amount of leaf primordia was detected in larger bulblets. Furthermore, the histological examination revealed that in L. candidum, as opposed to other lily species, there had been no real "phase change" in the meristem and that the phase change from juvenile to vegetative adult occurred at a much later stage in L. candidum than in other species.     

Effects of storage temperature and sucrose on bulblet growth, starch and protein contents in in vitro cultures of Hyacinthus orientalis

The scale segments of the bulblets of Hyacinthus orientalis L. cv. Anna Marie were examined to improve their growth and development with cold-pretreatment and sucrose. The cold-pretreated (4 °C for 4 months) segments showed higher growth and better development of the bulblets on medium without sucrose than ones stored at 20 °C. A rapid decrease in starch content of bulb pieces was found during the first 2 weeks in all cultures and thereafter the content decreased gradually. A scanning electron microscopic observation during the bulblet growth and development showed a gradual decreasing trend of the starch granules from 2 to 16 weeks of the cultures. SDS-PAGE electrophoresis revealed the presence of a characteristic polypeptide of approximately 45 kD, which is assumed to be a major storage protein in the bulblets.     

High frequency of shoot multiplication and bulblet formation of garlic in liquid cultures

In vitro shoot proliferation and bulblet production of garlic (Allium sativum L.) was studied in liquid cultures. Shoots grown in vitro were used as explants and were cultured in MS medium supplemented with 2% (w/v) sucrose and 0.5 mg l^–1 2-iP. Three culture methods (semi-solid, liquid-immersion and raft) were compared for shoot proliferation. Explants in liquid (immersion) culture exhibited an increased multiplication rate and fresh weight of shoots after 3 weeks of culture as compared with the other treatments. Bulblet formation and growth were studied in liquid medium with different concentrations of sucrose (2–13%). MS medium containing 11% (w/v) sucrose was optimal for bulblet development and bulblets developed in this medium within 9 weeks in culture. The highest multiplication rate was (135 bulblets/explant) found when explants were cultured in bulbing medium (MS medium containing 0.1 mg l^–1 NAA+11% (w/v) sucrose) supplemented with 10 M JA. Growth retardants CCC, B-9, ABA also promoted induction and growth of bulblets. Darkness promoted the bulblet induction and growth compared to light conditions (16-h photoperiod of 50 mol m^–2 s^–1). The dormancy of bulblets was broken by cold treatment at 4 °C for 8 weeks.     

Effect of low temperature on dormancy breaking and growth after planting in lily bulblets regenerated in vitro

Lilies regenerating on scale segments may develop dormancy in vitro depending on the culture conditions. The dormancy is broken by storage for several weeks at a low temperature (5 °C). The effect of the low temperature on sprouting, time of leaf emergence and further bulb growth was studied. Dormant and non-dormant bulblets were regenerated in vitro on bulb scale segments cultured at 20 °C or 15 °C, respectively. The low temperature not only affected the number of sprouted bulblets but also the time of emergence. The longer the cold storage, the faster and more uniform leaf emergence occurred. Both dormant and non-dormant bulblets grew faster after a low temperature treatment of six weeks. Thus, during dormancy breaking the tissue is prepared not only for sprouting but also for subsequent bulb growth. These processes are rather independent as low temperature stimulates growth in non-dormant bulblets whereas these bulblets sprout also without treatment at low temperature. Moreover, the hormone gibberellin induces rapid sprouting but has no influence on further bulb growth. Good growth in bulblets exposed to the low temperature coincided with production of an increased leaf weight. However, the relationship is not absolute as bulblets that were cold-treated for six weeks grew larger than bulblets cold-treated for four weeks but the formation of leaf biomass was similar. During storage at low temperature starch was hydrolyzed in the bulb scales and sugars accumulated. This indicates that during this period, preparation for later bulb growth involves mobilization of carbohydrate reserves which play a role in leaf growth and development of the photosynthetic apparatus. Starch hydrolysis proceeded in the outer scales after planting. Approximately six weeks later, the switch from source to sink took place in the bulblet, which became visible as a deposition of starch in the middle scales.     

Petal: a reliable explant for direct bulblet regeneration of endangered wild populations of <Emphasis Type="Italic">Fritillaria imperialis</Emphasis> L.

Wild populations of Fritillaria imperialis L. are facing extinction and need urgent conservation. This paper presents an efficient system for in vitro direct bulblet regeneration of these populations by petal culturing of flower buds. Petals at different developmental stages, green-closed flower bud (before nectar secretion) and red-closed flower bud (beginning of nectar secretion), were used as explants, and the effects of various proportions of cytokinin to auxin on direct bulblet regeneration pathway were evaluated. More explants switched on direct regeneration pathway in combination of auxins (0.6 mg l⁻¹ NAA + 0.4 mg l⁻¹ IAA) with higher level of cytokinin (1 mg l⁻¹ BAP). In contrast, auxins (0.6 mg l⁻¹ NAA + 0.4 mg l⁻¹ IAA) with lower level of cytokinin (0.1 mg l⁻¹ BAP) produced more bulblets per regenerated explant. In green-closed flower bud stage, direct bulblets regenerated from the end of petal where it was connected to the receptacle, while nectar secretion site was the place of bulblet formation in red-closed flower bud stage. In addition, genotype-dependency of direct bulblet regeneration pathway was investigated by using two different wild populations of Fritillaria imperialis. This plant regeneration procedure was applicable to different Fritillaria genotypes and regenerated bulblets were normal.