改进水稻花药培养方法的途径

用液面漂浮方式培养水稻花药,研究了培养基的蔗糖浓度、激素成分、附加秋水仙碱对花粉愈伤组织的诱导、分化及花粉植株倍性的影响。提出改进水稻花药培养方法的途径,其要点是:用液体培养基培养花药;在培养初期,调整培养条件以增加对等核分裂花粉数和提高染色体加倍频率;在多细胞团花粉突破花粉壁生长之前,将其从开裂的花药中分离出来进行单花粉固着培养形成愈伤组织,并使愈伤组织在生长的早期转向分化。     

小麦花药培养中花药密度效应的初步研究

在大麦花药培养中,花药密度对花粉愈伤组织产量有明显影响。欲得大量的愈伤组织,每毫升液体培养基上需接种60枚甚至更多的花药。利用条件培养基,可提高单位体积培养基上花药有效密度,进而提高大麦花粉愈伤组织的产量。在水稻花药培养中,花粉愈伤组织产量也有随花药密度而提高的趋势。     

大麦雄核发育的基因型效应及相关的RAPD标记

以具有不同花粉胚胎发生能力的两个大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅ Ｌ．）基因型为材料,对离体培养花药中不同类型小孢子频率的边续统计表明,两基因型的早期雄核发育过程无明显区别,最终出愈率（平均每个花药产生的愈伤组织块数）的显著差异是在小孢子形成多核花粉以后产生的。推测基因型之间胚胎发生能力的差异不受小孢子脱分化启动能力控制,而是由其多核花粉发育成愈伤组织的能力决定的,可能与细胞壁的形成有关。利用     

冬小麦的不同生育条件对其花粉植株诱导频率的影响

对三种不同生育条件的冬小麦的花药培养进行了比较,观察到它们对培养的反应有明显的和稳定的差异。经春化处理的冬小麦顶凌春播,5月中旬取花药做离体培养,比适时秋播,4月下旬进行离体培养的花粉愈伤组织和花粉植株的诱导频率提高约一倍;比冬天温室栽培,于2月下旬至3月上旬取花药做离体培养的愈伤组织诱导频率提高约二倍。1978年接种顶凌播种的28个杂交组合的冬小麦花药100％的组合诱导出了花粉愈伤组织,92.3％的组合分化出花粉绿苗。分析讨论了冬小麦顶凌播种诱导频率高的原因和它在冬小麦花粉单倍体育种研究中的应用     

甲基磺酸乙酯对水稻花药离体培养效应的研究

本文以粳稻“105”、籼稻“华03”和“鄂早6号”为实验材料,研究了不同浓度化学诱变剂EMS处理对离体培养水稻花药中花粉细胞脱分化与再分化和培养初期花药呼吸作用的影响。结果表明,低浓度EMS能显著提高花药愈伤组织诱导率,高浓度EMS则有明显的抑制作用。EMS对培养初期花药呼吸作用的影响与花药愈伤组织诱导率之间存在明显的平行关系。     

甲基磺酸乙酯对水稻花药离体培养效应的研究

本文以粳稻“105”、籼稻“华03"和“鄂早6号”为实验材料,研究了不同浓度化学诱变剂EMS处理对离体培养水稻花药中花粉细胞脱分化与再分化和培养初期花药呼吸作用的影响。结果表明,低浓度EMS能显著提高花药愈伤组织诱导率;高浓度EMS则有明显的抑制作用。EMS对培养初期花药呼吸作用的影响与花药愈伤组织诱导率之间存在明显的平行关系。     

药用植物组织培养的研究——Ⅲ、三分三愈伤组织的分化对莨菪碱、东莨菪碱含量的影响

从三分三种子萌发的根、茎、叶和花药、种皮诱导的愈伤组织,均含有莨菪碱、东莨菪碱。其含量和生长速度均以茎、叶愈伤组织为最高。根、茎、叶愈伤组织,在培养基中分别加入各种植物激素,培养时表明:BA(6-苄基嘌呤)促进芽的分化,NAA促进根的分化,2,4-D则抑制根的分化,GA_3(赤霉酸)影响很小。获得了三种愈伤组织只分化根不分化芽(加NAA 1毫克/升)和只分化芽不分化根(每升中加BA0.2毫克和2,4-D 0.2毫克)的良好结果。分化的这些芽或根的诱导频率,以茎、叶愈伤组织为高。分化的芽或根中均含有莨菪碱及东莨菪碱,但其含量相应地比它们的未分化的愈伤组织约低1～4倍。薄层层析结果表明,愈伤组织中有6种生物碱,茎、叶愈伤组织中,还有一种在紫外光下显蓝色荧光的物质,而分化的根或芽及花药愈伤组织中只有4种生物碱。看来,在离体培养下的三分三愈伤组织是有莨菪碱和东莨菪碱合成的全能性。这两种生物碱似乎并非最终的代谢产物,它们可能参与了器官建成中的代谢过程。文中还讨论了莨菪碱和东莨菪碱的合成部位问题。     

在低温预处理及花药培养期间高氯酸汞处理对大麦花药生产力的影响

在春大麦(sabarlis品种)幼穗低温预处理过程中,使用乙烯的吸收剂高氯酸汞溶液处理,明显提高花药培养中花药的诱导率,以及每个花药形成的愈伤组织数目。在花药培养中,此种处理仅增加花粉愈伤组织的形成,而对花药的诱导率影响不明显。     

影响小麦(Triticum vulgare)花粉植株诱导频率的几种因素

在离体培养条件下,对影响小麦花粉愈伤组织的诱导和分化的某些因素进行了对比试验;对部分组合的愈伤组织进行了染色体观察,其结果:1.做为碳源和调节渗透压的蔗糖,10%的浓度最适宜;2.酪蛋白水解物对花粉愈伤组织的形成,器官分化及细胞染色体的自然加倍有明显效果;3.花药接种前在3—5℃下处理48小时,对提高诱导频率有明显作用;4.较高的温度对愈伤组织的诱导及其分化能力有促进作用;5.遗传型的差异,对花粉植株的成功率起着一定作用。     

离体培养菠萝的分化

菠萝的各种器官(幼嫩的聚花果、花药、吸芽或裔芽的腋芽、小的冠芽和小的裔芽)培养在含有1—10毫克/升的 NAA(萘乙酸)和 BA(6—苄基腺嘌呤)的 MS 培养基上。花药的培养是采用含有四分体至成熟花粉各时期的材料,但均没有观察到形成花粉愈伤组织,偶而能形成花丝愈伤组织,     

LEVER ACTION ANTHERS AND THE FORCIBLE SHEDDING OF POLLEN IN TORENIA (SCROPHULARIACEAE)

The anthers of Torenia fournieri were found to shed pollen forcibly by lever action. Anther structure was modified for this function by a flangelike outgrowth of the lateral pollen sac wall forming a lever. When pressed, this lever causes an infolding of the thinner, subadjacent, pollen sac wall forcing pollen from the stomium. A force of 1–1.5 g pressing against the four levers of an anther pair resulted in the forcible shedding of 2,000–3,000 pollen grains in two parallel rows. During the 2-day anthesis the flowers shift from functioning as males to hermaphroditic outcrossers, and yet only have a pollen:ovule ratio of 98.6, a ratio more indicative of facultative autogamy. The outermost pair of anthers functions on the first day of anthesis, while the second, inner pair functions on both days. In each flower, the 2-day anther pair produces approximately twice as many pollen grains as the 1 -day anther pair, a pollen production highly correlated with the length of their functional lives. This difference in pollen production is apparent in the larger size of the 2-day anthers, a size difference that first appears with the initiation of the anther primordia.     

EMBRYOGENIC DETERMINATION AND RIBONUCLEIC ACID SYNTHESIS IN POLLEN GRAINS OF HYOSCYAMUS NIGER (HENBANE)

³H-uridine administered as a one- or two-hour pulse to embryogenic pollen grains of freshly excised anthers of Hyoscyamus niger (henbane) was autoradiographically localized in embryoids formed during a subsequent chase. Although continuous incubation of anthers in actinomycin D inhibited embryogenesis, a small percentage of potentially embryogenic pollen escaped inhibition if anthers were grown for at least one hour in the basal medium before actinomycin treatment. The results imply that certain pollen grains become embryogenically determined immediately after culture of the anther and that this is accompanied by the synthesis of ribonucleic acid.     

Studies on Induction of Pollen Plantlets from the Anther Cultures of Lily

Anthers with the filament of lily (Lilium davidii var. Willmottiae (Wilson) Roffill) were cultured on modified MS medium. Supplemented with different concentrations and compatible ratios of growth hormones (Z 2 mg/L,or 2,4-D 2 mg/L + KT 2mg/L, or 2,4-D 4mg/L+ 6 BA 2 mg/L). At this time the pollen grains in the anthers were at the late uninueleate stage. Anther cultures were incubated at 25—27 ℃, and illuminated with daylight of about 800–1200 lx. After 30 days, the calli or embryoids were produced from anthers. The frequency of the calli or embryoids induction was 8.89%. After transfer eventually to the differentiation medium, these calli or embryoids developed into plantlets in 70 days. Among the root tips of regenerated plantlets haploid, diploid and aneuploid cells were found, but the haploid cells were produced in about 86.4% of the root tips. It is quite evident that haploid plantlets are derived from the pollen grains.     

低温预处理影响水稻花药培养效率的机理初探

低温预处理延缓药壁中层和绒毡层的降解,促进表皮层和药室内壁层的发育,延缓花药过氧化物酶同工酶活性的增强。处理期间花药可溶性蛋白质、淀粉酶同工酶潜带发生明显变化。处理期间花药的~3H-TdR渗入和花粉的发育、分裂,表明花粉存在合成和充实活动。绒毡层和花粉间存在囊泡,表皮层和药室内壁层之间存在多泡体的穿壁运动,说明低温处理中药壁向花粉输送雄核发育所需的物质。在进入正常培养初期,经过低温处理的花药药壁仍有表皮层和药室内壁层的发育,多细胞花粉出现提早、数量增加,花粉退化延缓。而未经处理的花药药壁各层均迅速降解,花粉大量退化。     

石刁柏花粉植株诱导及其起源鉴定的研究

采用高渗蔗糖溶液预处理石刁柏花药可以显著抑制花药体细胞分裂和提高花粉愈伤组织诱导率。愈伤组织在转入含低浓度激素的培养基中分化得到了花粉植株。其中单倍体、二倍体、四倍体和非整洁体分别占４．３％、６４．５％、１７．２％和１４．０％。单倍体的频率随愈伤组织培养时间延长而下降,石刁柏幼茎中莽草酸脱氢酶同工酶的多态性表现稳定,用其作为遗传标记结合细胞学方法可以鉴定花粉植株的起源。     

丹参雄性不育系Sh-B的鉴定与花粉发育过程的解剖学研究

在显微水平上对新发现的丹参雄性不育系Sh-B花药发育过程进行了解剖学观察,并对其花粉活力和结实率进行了鉴定。结果显示:根据花器官及花药的形态、大小以及花丝的长度,可以将Sh-B不育株分为3个不育类型,即Sh-B1、Sh-B2和Sh-B3。这3种不育类型均属于雄性不育,其花丝不到正常可育株的1/2,花药干瘪而瘦小,内无花粉粒或花粉无活力;其根、茎、叶以及种子形态结构与正常可育植株基本相似。产生雄性不育的主要原因有:花粉囊药室内壁纤维层加厚,影响花药壁开裂;小孢子母细胞周围不产生胼胝质或产生的胼胝质很少;绒毡层细胞延迟解体;花粉粒畸形。在其花药发育的小孢子母细胞时期、四分体形成前期、单核期、双核期均可能产生雄性不育的小孢子或花粉粒。     

THE CYTOLOGY OF POLLEN ABORTION IN S CYTOPLASMIC MALE-STERILE CORN ANTHERS

The anther development of the S male-sterile cytoplasm and the fertile maintainer (N) cytoplasm versions of corn inbred W182BN and the restored S cytoplasm version of inbred NY821LERf was studied by light and electron microscopy and compared to pollen abortion in the C and T types of male-sterile cytoplasms. The S anthers did not deviate from the non-male sterile (N) anthers until a very late stage of pollen development. Tapetal cells developed and disappeared normally in the S version which differentiates this cytoplasm from the C and T types. Although some modified membranous structures were seen in a higher frequency in the large vacuole of the sterile S pollen than in the N and restored S counterparts, the mitochondria and other organelles in the S pollen appeared normal up to the time of pollen abortion. Pollen abortion in the S cytoplasm did not occur until the developing pollen was nearly mature. At this time the pollen grains disintegrated abruptly but other anther tissues appeared unaltered. The male sterility of S plants appeared to be determined by the pollen itself without external influence from the tapetum.     

Anther Factor(s) in Barley Anther Culture

The effects of anther tissues were studied systematically on microspores forming multicellular units and furthermore on pollen callus formation in the anther culture of Hordeum vulgare (cv. Sabarlis). Anther productivity was found to be greatly enhanced by use of medium previously conditioned by anthers. In 15 experiments observed, anthers produced 26 times on average more calli in the conditioned medium than in control, in a few cases, even more than 80 times more calli formed. According to this, the authors supposed that cultured anthers released some components, anther factor (s) (AF), which is important to androgenesis in the culture. To achieve high yields of callus, culture was restricted to anthers which had been subpected to cold pretreatment. The temperature stress could not be replaced by the AF. However, for conditioning medium, anthers at binuclear stage were found to be more effective than the test anthers either with or without the pretreatment. Anthers from other 8 barley varieties were also effective for conditioning, as the difference of anther productivity still existed in the culture with conditioned medium between various genotypes tested. Anther response and callus yield were increased in both the culture of anthers at mid and late-uninuclear stage by use of conditioned medium. AF interacted synergistically with m-inositol. Cytological observation showed that AF increased apparently the formation of MPGs, while m-inositol mainly stimulated callus formation from MPGs. To some extent, the effect of exogenous hormone(s) could be replaced by AF. The anther response and pollen callus yield could be much enhanced by increasing anther inoculation density, which also raised the AF level in the culture. Thus, by use of the temperature stress prior to anther culture and culture of test anthers in conditioned medium with m-inositol, or at higher inoculation density, a very high production of pollen callus could be obtained in barley anther culture. For meeting the more specialized requirements of less responsive species or genopypes, the principles given here may be provide some basic information.     

低温预处理影响籼稻花药培养效率的研究

低温预处理使籼稻花药在培养初期退化花粉百分率低于对照;多核花粉百分率明显高于对照。经过低温预处理的花药来见新的可溶性蛋白谱带出现;淀粉酶同工酶谱带少和弱于对照,过氧化物酶同工酶中碱性区活性初期弱而后期强于对照。低温处理使药壁组织衰退进程有所延缓,但未能制止衰退,也未出现如粳稻中所见的药壁细胞充实发育的现象。低温预处理只能稍微延缓籼稻药壁组织的衰退进程,有限度地促进籼稻花药培养效率。