小麦成熟胚愈伤组织诱导及分化研究

以2个小麦品种成熟胚为外植体进行离体培养,研究了不同预处理、不同2,4-D浓度及与KT组合、不同蔗糖浓度等因素对愈伤组织诱导及分化的影响。结果表明:4℃低温预处理可提高愈伤组织的出愈率及再生苗率,2个材料的出愈率及再生苗率均达到90%和30%以上;在不同预处理条件下,2,4-D浓度对出愈率及再生苗率的影响与基因型有关,2,4-D浓度为1~2 mg/L更有利于愈伤组织诱导及分化;附加KT能缓解高浓度2,4-D对再生苗率的抑制作用,而对于在1、2 mg/L 2,4-D的培养基中附加KT则不表现这种作用;蔗糖浓度则在30 g/L条件下更有利于愈伤组织诱导。因此通过4℃低温预处理,在MS基本培养基中附加1~2mg/L 2,4-D及30 g/L蔗糖亦可促进小麦成熟胚愈伤组织的诱导和分化。     

小麦未成熟胚诱生大量绿苗的研究初报

根据6个春麦品种及4个冬麦品种的试验结果,提高诱导小麦未成熟胚绿苗率的关键是:(1)掌握最适宜的接种时间,并要求幼胚的盾状组织朝上放置。(2)诱导愈伤组织的培养基(A)的最佳配方为MS培养基,3％蔗糖,245-T 2mg/J,IAA0.4mg/J,激动素0.2 mg/J,琼脂0.75％,pH5.7。主要改动是以245-T代替2,4-D,并提高IAA的浓度。新方法的愈伤组织诱导率高达85—100％,以幼胚为计算基础的绿苗诱导率为84—99％,每个萌动的幼胚一般可产生健壮的绿苗16—20株。     

Water Content, Raffinose, and Dehydrins in the Induction of Desiccation Tolerance in Immature Wheat Embryos

Desiccation tolerance is initiated in wheat (Triticum aestivum L.) embryos in planta at 22 to 24 d after anthesis, at the time that the embryo water content has decreased from about 73% fresh weight (2.7 g water/g dry weight) to about 65% fresh weight (1.8 g water/g dry weight). To determine if desiccation tolerance is fully induced by the loss of a relatively small amount of water, detached wheat grains were treated to reduce the embryo water content by just a small amount to approximately 69% (2.2 g water/g dry weight). After 24 h of such incipient water loss, subsequently excised embryos were able to withstand severe desiccation, whereas those embryos that had not previously lost water could not. Therefore, a relatively small decrease in water content for only 24 h acts as the signal for the development of desiccation tolerance. Embryos that were induced into tolerance by a 24-h water loss had no detectable raffinose. The oligosaccharide accumulated at later times even in embryos of detached grains that had not become desiccation tolerant, although tolerant embryos (i.e. those that previously had lost some water) contained larger amounts of the carbohydrate. It is concluded that desiccation tolerance and the occurrence of raffinose are not correlated. Immunodetected dehydrins accumulated in embryos in planta as desiccation tolerance developed. Detachment of grains induced the appearance of dehydrins at an earlier age, even in embryos that had not been made desiccation tolerant by incipient drying. It is concluded that a small reduction in water content induces desiccation tolerance by initiating changes in which dehydrins might participate but not by their interaction with raffinose.     

ABA Levels and Sensitivity in Developing Wheat Embryos of Sprouting Resistant and Susceptible Cultivars

A sprouting-resistant and a sprouting-susceptible wheat cultivar were utilized to examine the role of ABA levels and sensitivity responses in wheat embryonic germination. Endogenous embryonic ABA levels were measured in both cultivars throughout grain maturation utilizing a new and sensitive ABA immunoassay. Embryonic ABA levels of each cultivar were similar with the sprouting-susceptible cultivar having about a 25% lower ABA level than that of the sprouting-resistant cultivar. Larger differences between the cultivars were noted in sensitivity to ABA, as measured by capability of ABA to block embryonic germination. ABA inhibited embryonic germination much more effectively in the sprouting-resistant cultivar.     

The Soluble Nucleotide Content of Germinating Wheat Embryos

Estimates are given of the amounts of adenine, uracil, guanine,and cytosine present in the soluble nucleotides of wheat embryosduring germination in the dark. After 48 hours at 25° Cthe nucleotide content per gramme of dry tissue reaches a maximumlevel when the content of bases is approximately 3.5 µmolesof uracil, 2.8 µmoles of adenine, 0.6 µmoles ofguanine, and 0.5 µmoles of cytosine. The soluble nucleotidecontent of embryos growing at 0 to 5° C is lower than thatof embryos of the same dry weight grown at 25° C. Initiallythe amount of adenine present is greater than the amount ofuracil but after 20 hours of germination at 25° C uracilbecomes the predominant base in the soluble nucleotide fraction.Ion-exchange resin chromatography was used to separate the chiefsoluble nucleotides present in the extract of embryos grownfor four days at 25° C. Uridine diphosphate glucose accountsfor the major part of the uracil and adenosine monophosphatefor most of the adenine. Nicotinamide-adenine dinucleotide wasdetected and identified and also a little uridine 5'-mono-phosphate.The possibility is discussed that most of the adenosine monophosphateis produced by degradation of reduced nicotinamide-adenine dinucleotidephosphate.     

南方红豆杉试管微芽诱导培养及其紫杉醇类化合物的积累

以南方红豆杉幼茎段和茎尖为外植体,以wPM为基本培养基,单一添加不同浓度的KT、TIBA、ZT和6-BA进行芽诱导培养。结果表明：各自添加0．4mg·L-1KT、3．0mg·-1TIBA、0．1mg·L-1 ZT和0．01mg·L-1 6-BA时的芽诱导率最高。培养50d的试管微芽中紫杉醇和10一去乙酰基巴卡亭III（10．DABIII）的积累量比天然南方红豆杉嫩枝高2倍以上,试管微芽中10一DABIII比紫杉醇高,且二者均随芽诱导率的增加而增加,当芽诱导率达最高时亦达到最高。TIBA所诱导芽短而粗壮。     

Development and Floret Survival in Wheat Ears With Supernumerary Spikelets

In the supernumerary spikelet wheat, AUS159I0, the supernumeraryspikelet primordia appeared just after the ear reached the terminalspikelet stage. Appearance of the primordia of the multiplesessile spikelets preceded that of indeterminate rachilla spikelets.Supernumerary spikelets had a lower number of potentially fertileflorets per spikelet than normal (non-supernumerary) spikeletsin the ear and thus a smaller number of grains per spikelet.Mean weight per grain in the supernumerary spikelet wheat waslower than in the cultivar, Meering, without supernumerary spikelets.Total grain number in the supernumerary spikelet ear was greaterthan in the normal ear despite the lower spikelet fertilityin the former. Within the supernumerary spikelet ear the multiplesessile spikelets had a higher number of grains per spikeletand mean weight per grain than the indeterminate rachilla spikelets.It appears possible to improve the productivity of the supernumeraryspikelet ear by breeding for reduced expression of the indeterminaterachilla spikelets. Wheat, ear development, floret survival, supernumerary spikelets, grain number     

小麦幼芽水分胁迫诱导蛋白的特征

水分胁迫（－1．2MPaPEG-6000）处理萌动的小麦种子,24h后诱导小麦幼芽产生41．5kD蛋白,其含量随着胁迫时间延长明显增加,48h时含量最高,到72h后不再变化。复水后,该蛋白消失;再胁迫48h时则又出现,其含量与处理24h时相当。41．5kD诱导蛋白主要位于细胞器膜上,细胞质中几乎不存在。41．5kD蛋白主要溶于10％NaCl提取液中,其等电点为pl5．65。该蛋白的氨基酸组成中,脯氨酸含量最高,其次为丙氨酸、天冬氨酸、谷氨酸、甘氨酸,没有发现半胱氨酸和组氨酸。     

Isoelectrofocusing Electrophoretic Analysis on Soluble Protein of Germinated Embryos and Young Shoots from Different Cytoplasmic Male Sterile Lines and Their Maintainers in Wheat

The soluble proteins of germinated embryos and young shoots in Wheat were studied by means of isoelectrofocusing (IEF) electrophoresis. The materials used were three species of cytoplasmic male-sterile (CMS) lines (Type E, T and A) and their maintainers. The protein quantity of pI 4. 9 in male-fertile materials is higher than the corresponding male-sterile ones; the protein of pI 6.85 is probably the result of gene expression in Timopheevi cytoplasm; the protein of pI 7.6 is a typical band of Jinfeng male-sterile line. Evident differences on soluble protein chromatograms of IEF electrophoresis were observed both in germinated embryos and young shoots which were from different kinds of CMS line, these differences could be used as indexes identifying various CMS lines.     

The Phospholipid Composition of Nuclear Subfractions from Germinating Wheat Embryos

The content of phospholipids in chromatin, nuclear matrix, and nuclear membrane from wheat (Triticum aestivum L.) embryos was studied. Subfractions of intact nuclei from dry embryos were shown to differ in the content and composition of particular phospholipids. Embryo germination resulted in the redistribution of phospholipid between nuclear subfractions. A functional role of structural changes in the nuclear membrane due to this phospholipid redistribution is discussed. It is supposed that these rearrangements change nuclear membrane permeability and its surface charge.     

Influence of Vernalization and Photoperiod on Supernumerary Spikelet Expression in Wheat

Two tetraploid (Triticum turgidum L.emend gr. turgidum and gr.durum) and five hexaploid wheats (Triticum x aestivum L. emendgr. aestivum) with reported tendencies for ‘branched heads’(supernurnerary spikelets) exhibited variation in its expressionunder different vernalization photoperiod and temperature regimes. Two main types of supernumerary spikelets were identified, multiplesessile spikelets (MSS) with two or more complete spikeletsat a rachis node and indeterminate rachilla spikelets (IRS)with two to 13 spikelets on an extended rachilla. The degree of supernumerary spikelet expression in wheats withvernalization response differed from those without. Short photoperiods(9–14 h) both outdoors and in a glasshouse environment,were more conducive to supernumerary spikelet expression than24 h photoperiod in both environments. The 24 h photoperiodglasshouse environment (higher mean temperatures) was leastconducive to its expression except in lines with a strong vernalizationresponse. The high stability of supernumerary spikelet expression in certaingenotypes in the different environments indicated the feasibilityof incorporating this character in breeding and selecting commercialwheats to increase grain number per head. Triticum, wheat, ear-branching, supernumerary spikelets, vernalization, photoperiod, temperature     

Studies on the Formation of Roots and Shoots in Wheat Callus Cultures

Callus tissues were initiated from root, embryo and inflorescenceexplants of wheat. These callus cultures were used to studythe formation of roots and shoots in the absence and presenceof selected plant hormones. On a basal medium alone, only newly-initiatedembryo callus formed both roots and shoots while root callusonly formed roots. Inflorescence callus showed no signs of differentiation.The regenerative capacity of root and embryo callus tissueson medium lacking hormones decreased with increasing periodsof culture. Calluses which failed to differentiate in the absenceof hormones were selected for studies on hormone-mediated differentiation.NAA (1 mg 1^–1) was effective in inducing roots from allcalluses irrespective of their origin or age. In contrast, shootformation was elicited by incubating newly-formed callus onbasal medium supplemented with kinetin (5 mg 1^–1) andNAA (1 mg 1^–1) but rapidly decreased with longer periodsof culture. The differences observed in differentiation of thecallus in the absence and presence of hormones is discussed.     

The Onset of Germination Ability in Developing Wheat

The ability of primary wheat grains (cv. Sappo) to germinatewas determined at different times from anthesis. Fresh, prematurelyharvested grains germinated best at low temperatures (<10°C). The temperature range over which appreciable germinationtook place broadened and the maximum percentage germinationachieved increased with lengthening time between anthesis andharvest. Chilling and exogenous applications of GA₃ each affectedthe rate of germination and the maximum percentage germinationof grains in their own ways. The results are discussed in relationto the identification of ‘dormancy’ in wheat grains.     

铁皮石斛茎段诱导丛生芽的研究

以铁皮石斛Dendrobium candidum Wall.ex Lindl.的茎段作外植体,比较不同的培养基、激素等因素对茎段分化丛生芽的影响,以及不同浓度的香蕉汁和活性炭对试管苗生根的影响。结果表明：1／2MS较好,BA作用优于KT、ZT,最适浓度为2．0mg/L;丛生芽培养于1／2MS 10％香蕉汁 0．5％AC的培养基上,生根效果最好。     

The Negative Influence of High-Glucose Ambience on Neurogenesis in Developing Quail Embryos

Gestational diabetes is defined as glucose intolerance during pregnancy and it is presented as high blood glucose levels during the onset pregnancy. This condition has an adverse impact on fetal development but the mechanism involved is still not fully understood. In this study, we investigated the effects of high glucose on the developing quail embryo, especially its impact on the development of the nervous system. We established that high glucose altered the central nervous system mophologically, such that neural tube defects (NTDs) developed. In addition, we found that high glucose impaired nerve differentiation at dorsal root ganglia and in the developing limb buds, as revealed by neurofilament (NF) immunofluorescent staining. The dorsal root ganglia are normally derived from neural crest cells (NCCs), so we examine the delamination of NCCs from dorsal side of the neural tube. We established that high glucose was detrimental to the NCCs, in vivo and in vitro. High glucose also negatively affected neural differentiation by reducing the number and length of neurites emanating from neurons in culture. We established that high glucose exposure caused an increase in reactive oxidative species (ROS) generation by primary cultured neurons. We hypothesized that excess ROS was the factor responsible for impairing neuron development and differentiation. We provided evidence for our hypothesis by showing that the addition of vitamin C (a powerful antioxidant) could rescue the damaging effects of high glucose on cultured neurons.     

Branching Shoots and Spikes from Lateral Meristems in Bread Wheat

Wheat grain yield consists of three components: spikes per plant, grains per spike (i.e. head or ear), and grain weight; and the grains per spike can be dissected into two subcomponents: spikelets per spike and grains per spikelet. An increase in any of these components will directly contribute to grain yield. Wheat morphology biology tells that a wheat plant has no lateral meristem that forms any branching shoot or spike. In this study, we report two novel shoot and spike traits that were produced from lateral meristems in bread wheat. One is supernumerary shoot that was developed from an axillary bud at the axil of leaves on the elongated internodes of the main stem. The other is supernumerary spike that was generated from a spikelet meristem on a spike. In addition, supernumerary spikelets were generated on the same rachis node of the spike in the plant that had supernumerary shoot and spikes. All of these supernumerary shoots/spikes/spikelets found in the super wheat plants produced normal fertility and seeds, displaying huge yield potential in bread wheat.