Tolerance analysis for incident angle of volume phase holographic grating used in optical channel performance monitor

Transmission Volume Phase Holographic Grating (VPHG) is adopted as spectral element in the real-time Optical Channel Performance Monitor (OCPM), which is in dire need in the Dense Wavelength -division-multiplexing(DATDM) system. And the tolerance of incident angle, which can be fully determined by two angles: 6 and (p, is finally inferred in this paper. Commonly, the default setting is that the incident plane is perpendicular to the fringes when the incident angle is mentioned. Now the situation out of the vertical is discussed. By combining the theoretic analysis of VPHG with its use in OCPM and changing 6 and (0 precisely in the computation and experiment, the two physical quantities which can fully specify the performance of VPHG the diffraction efficiency and the resolution, are analyzed. The results show that the diffraction efficiency varies greatly with the change of 6 or (p. But from the view of the whole C-band, only the peak diffraction efficiency drifts to another wavelength. As for the resolution, it deteriorates more rapidly than diffraction efficiency with the change of (p, while more slowly with the change of theta. Only if \phi\less than or equal to+/-1degrees and alpha(B) -0.5 less than or equal to theta less than or equal to alpha(B) + 0.5, the performance of the VPHG would be good enough to be used in OCPM system.

青杨组(Populus section Tacamahaca Spach)不同种对增强UV-B的反应差异

人类向大气中排放的大量氮氧化合物和氟氯烃类化合物(CFC’s)引起臭氧分子的分解，导致到达地球表面的紫外辐射增加，特别是UV-B辐射增强。本项目以青杨组杨树为模式植物，从形态和生理方面研究了来自不同UV-B背景下的康定杨与青杨在增强UV-B下的反应及其反应差异,并探讨了干旱、施肥对它们抗UV-B能力的影响。杨树具有分布广、适应性强、在生态环境治理和解决木材短缺方面均占有重要位置，研究成果可为生态系统的恢复与重建提供理论依据和科学指导。主要研究结果有以下： 1. 在温室中经过增强UV-B处理，杨树的外部形态及生理活动受到了一定程度的影响。增强UV-B导致康定杨、青杨的生物量、叶面积及节间长度降低，叶片增厚，SOD活性升高，膜伤害增加，而对叶片数目、R/S、叶绿素A、叶绿素B及整个叶绿素含量没有影响。两种杨树对UV-B胁迫的响应存在差异：在增强UV-B条件下，青杨的植株高度、生物量、叶面积、脯氨酸含量、长期用水效率受到的影响大于康定杨，相比而言，康定杨在比叶面积、叶片厚度、可溶性糖含量、UV-B吸收物质的含量及SOD和GPX活性方面增加的程度大于青杨。这些区别说明，来自于高海拔的康定杨比来自于低海拔的青杨对增强UV-B 具有更强的耐性。我们认为二者在叶片厚度、比叶面积、UV-B吸收物质含量及SOD、GPX活性差异是导致对增强UV-B耐性不同的原因。 2. 干旱与增强UV-B对杨树的生长和生理特性均产生了影响，而且两种胁迫共同作用时干旱表现减弱或加剧了UV-B对杨树某些形态和生理特性的影响。 据试验结果，干旱显著地降低了杨树的株高、叶片数目、叶面积，增加了叶片厚度，促进ABA的积累，提高了CAT活性。对于干旱，两种杨树之间也表现出了一定的差异性。可溶性蛋白质和脯氨酸在青杨叶片中得到显著积累，而在康定杨中没有变化。此外，CAT、长期用水效率在康定杨中受到的影响更加明显。长期用水效率的不同变化趋势说明两种杨树对水分胁迫采用了不同的用水策略，康定杨采用的是节水用水策略，提高用水效率，而青杨采用的是耗水的用水策略。根据干旱对叶面积、脯氨酸、ABA含量、CAT活性及长期用水效率等方面的影响，我们认为来自高海拔地区的康定杨比来自低海拔的青杨有更大的耐旱性，这是对生长环境长期适应的结果。在高海拔地区，因霜冻常带来土壤水分不可利用，降低了根系对水分的吸收，树木容易受到的生理性干旱。另外，高海拔的地区低的气温使植株对严寒有较强的耐性，减少了水分的需要。 生长于增强UV-B下的康定杨和青杨植株表现为高度降低，叶面积缩小，比叶面积增加；叶片栅栏组织、海绵组织均受到增强UV-B的影响，其厚度的增加导致整个叶片变厚。增强UV-B还显著提高了杨树的APX活性、UV-B吸收物质含量，而对叶片数目、ABA、可溶性蛋白质含量及CAT活性没有产生影响。试验中也观察到了两种杨树对增强UV-B响应的差异：与康定杨相比，在增强UV-B下青杨株高、叶面积降低的程度更大一些，SOD活性显著提高。另外UV-B吸收物质受到的影响不同。根据这些差别，高海拔的康定杨(3500 m)比来自低海拔的青杨(1500 m)增强UV-B有较强的耐性。 与水分充足情况下UV-B对植株的影响相比，干旱对杨树抗增强UV-B产生了一定的影响，表现为加剧或减弱UV-B对植物的影响，但这种影响与形态、生理指标有关。当干旱与增强UV-B共同作用时，杨树植株的株高、叶面积进一步降低、叶片进一步增厚。就脯氨酸的积累的而言，在没有水分胁迫时，增强UV-B促使它显著增加，而在干旱处理下这种效果变得不明显。干旱对增强UV-B的影响还与杨树的种类有一定的关系。在康定杨中，干旱减弱了增强UV-B对栅栏组织与海绵组织的影响，且在植株高度、叶面积上表现出累加效应，而在CAT上交互作用显著；但在青杨中干旱则加剧增强UV-B对栅栏组织与海绵组织的影响，在植株高度、叶面积及比叶面积上表现出显著的交互作用。据碳同素分析，在水分充足的条件下，无论是康定杨，还是青杨，增强UV-B均导致其长期用水效率的提高，然而当两种胁迫共同作用时，长期用水效率则表现出差异，在青杨中，长期用水效率得到进一步增高，而康定杨中干旱的效应被增强UV-B所减轻。 3. 田间试验表明，杨树的生长、生理特征都受到养分和增强UV-B的影响。施肥对杨树的影响表现为：提高了叶面积、生物量及SOD的活性，降低了抗坏血酸含量。对于施肥作用，两种杨树的反应也有区别：在康定杨中施肥显著增加了的叶片长度、宽度及光合色素的含量，降低了净光合速率、气孔导度及胞间CO2浓度；在青杨中，则SOD、GPX、APX活性表现增加。从试验看出，施肥对来自于高海拔地区的康定杨(3500 m)的影响较大，对来自低海拔的青杨(1500 m)影响较小，这与它们对原产地的生境适应有一定关系。在康定杨生长的高海拔地区，低温度和湿度不能为地上凋落物或土壤中的根分解提供理想的条件，造成当地土壤的低养分状况，所以当肥料施用以后，效果显著。 经过增强UV-B处理，杨树叶片中UV-B吸收物质含量、GPX的活性得到提高，而脯氨酸、丙二醛、可溶性蛋白质、叶绿素及类胡萝卜素含量没有受到影响。对于增强UV-B两种杨树受到的影响也有所不同：在青杨中增强UV-B导致叶面积缩小，生物量、净光合速率降低，APX的活性及长期用水效率的提高，而对康定杨的这些指标没有产生显著影响，相反抗氧化酶的活性明显高于青杨。这些差异性是由于两种杨树对原产地不同UV-B背景的长期适应结果。康定杨长期生长在较高UV-B环境中，对UV-B有较强的耐性。而青杨适应于较低的UV-B环境，对增强UV-B较为敏感。 试验中施肥也影响了植株对增强UV-B的反应，不过这种影响与杨树的种类及测定指标有一定的相关性。例如，在缺肥的情况下，青杨的长期用水效率和康定杨的叶绿素含量都受到增强UV-B的显著影响，而施肥以后这种影响变得不显著。在缺肥的条件下，GPX、APX在青杨中的活性、GPX在康定杨中的活性对增加UV-B反应不敏感；而施肥以后则变化显著，同样胞间CO2浓度在康定杨也有类似的变化。 For past decades, Ultraviolet radiation, especially UV-B reaching the Earth’s surface increased because of depletion of ozone layer resulted from emission of NxO and CFC’s from human activities. In this experiment, different species of Populus section Tacamahaca Spach from different UV-B background were selected as a model plant to assess the effects of enhanced UV-B radiation. Morphological and physiological traits induced by enhanced UV-B were observed and the different responses between P. kangdingensis and P. cathayana were discussed, furthermore the influences of drought and fertilizer on responses induced by enhanced UV-B were studied. Since poplars play an important role in lumber supply, and are important component of ecosystems due to their fast growth and wide adaptation, the study could provide a strong theoretical evidence and scientific direction for the afforestation, and rehabilitation of ecosystem. The results are as follows: 1. The experiment conducted in a greenhouse indicated that morphological and physiological traits of two poplars were affected by enhanced UV-B radiation. Enhanced UV-B radiation not only reduced biomass, leave area and internode length, but also increased leaf thickness and SOD activity as well as MDA concentration and electrolyte rate. However, no significant changes in leaf numbers, root shoot ratio, and total chlorophyll and chlorophyll component were observed. There were different responses to enhanced UV-B radiation between two species. Compared with P. kangdingensis, cuttings of P. cathayana, exhibited lower height increment and smaller leaf area. In addition, there were significant differences in free proline, soluble protein, and UV-B absorbing compounds, and the activity of SOD and GPX, long-term WUE between them. Differences in plant height, biomass, leaf area, free proline concentration, and long-termed WUE showed that P. cathayana were more affected by enhanced UV-B radiation than P. kangdingensis. In contrast, more increase of specific leaf mass, leaf thickness, and soluble sugar, and UV-B absorbing compounds, and activity of SOD and GPX were observed in P. kangdingensis. According to these results, we suggested that P. kangdingensis from high elevation, which adapted to higher UV-B environments, had more tolerance to enhanced UV-B than P. cathayana from low elevation, which adapted to lower UV-B environment. We believe it was the difference of leaf thickness, specific leaf mass, and UV-B absorbing compounds as well as the activity of SOD and GPX resulted in lower adaptation of P. cathayana to enhanced UV-B radiation. 2. Growth and physiological traits of two poplars were affected by both drought and enhanced UV-B radiation. Moreover, it was observed that when two stresses applied together drought could exacerbate UV-B effects or decrease sensitivity to UV-B. In the experiment, drought significantly decreased plant height, leaf numbers, leaf area, and increased leaf thickness, and ABA, and CAT activity of two poplars. There were significant interspecific differences to drought stress. Exposed to drought, soluble protein and proline concentration were increased in P. cathayana but not in P. kangdingensis. However, more changes in CAT and long-term WUE were observed in kangdingensis. Different change in long-term WUE suggests that two poplars adapted different water-use strategies. P. kangdingensis employ a conservative water-use strategy, whereas P. cathayana employ a prodigal water-use strategy. Based on the differences in leaf area, accumulation of free proline and ABA, CAT activity as well as long-term WUE, we believed that P. kangdingensis from high elevation had a greater tolerance to drought than P. cathayana from low elevation，which is the result of adaptation to local environment. In high elevation area, trees are prone to suffer from physiological drought because of un-movable water caused by frost. Besides lower temperature enable the plants had greater adaptability to frost as a results the requirement of water is reduced Enhanced UV-B radiation decreased shoots height, leaf area, and increased specific leaf mass and thickness of palisade and sponge layer as well as APX activity and UV-B absorbing compounds in both species. Whereas, leaf numbers, ABA content, soluble protein and CAT activity showed no differences to enhanced UV-B radiation. Interspecific differences were also observed. Compared with P. kangdingensis, P. cathayana showed lower shoot height and smaller leaf area, higher SOD activity. Besides, variation in UV-B absorbing compounds was found. These differences suggested that P. kangdingensis from high elevation (3500 m) was more tolerant to enhanced UV-B radiation than P. cathayana from low elevation (1500 m). Compared with morphological and physiological changes induced by enhanced UV-B radiation under well-watered conditions, drought exacerbated or decreased these changes. However, these effects vary with parameters measured. When two stresses applied together, shoot height and leaf area further decreased while leaf thickness further increased. Under well-watered conditions, enhanced UV-B radiation significantly increased proline content, but such effect was not observed under drought conditions. The effect of drought on enhanced UV-B radiation was related to species. For example, drought reduced the effects of enhanced UV-B radiation on palisade parenchyma and sponge mesophyll in P. kangdingensis, and additive effects in shoot height and leaf area and interactive effect CAT activity were observed. In contrast, for P. cathayana drought significantly exacerbated the effects of enhanced UV-B radiation on palisade parenchyma and sponge mesophyll; there were noticeable interaction in shoot height, leaf area and specific leaf mass. As far as long-term WUE is concerned, it was increased by enhanced UV-B radiation under well-watered conditions in both species. While different effect was observed between two species in combination of two stresses. Long-term water use efficiency was further increased in P. cathayana whereas the effect was less significant in P. kangdingensis. 3. The field experiment showed that growth and physiological traits of poplars were affected by nutrition and enhanced UV-B radiation. Fertilization significantly increased leaf area, biomass and SOD activity, reduced Ascorbic acid concentration. There was interspecific difference in response to fertilization. For P. kangdingensis, fertilization significantly increased leaf width, leaf length and photosynthetic pigments content while net photosynthetic rate and stomatal conductance, intercellular CO2 concentration were significantly decreased. However, for P. cathayana, these parameters were unaffected except the increase of SOD, GPX and APX activity. From above, it could concluded that P. kangdingensis from high elevation was more affected by fertilization than P. cathayana, This difference was due to adaptation to local environment., The low temperature and moisture where P. kangdingensis was collected can not provided optimum to decompose roots and litter fall as a result the nutrition in soil was poor. Exposed to enhanced UV-B radiation, for both species UV-B absorbing compounds and GPX activity were significantly increased while proline, MDA, soluble protein, chlorophyll, carotenoids were not affected. Different responses were also observed between the two species. Enhanced UV-B radiation caused significant decreases in leaf area, biomass, net photosynthetic rate and increase in APX activity and long-term WUE in P. cathayana but not in P. kangdingensis. In addition, activity in antioxidant enzymes was much higher in P. kangdingensis than in P. cathayana. In the experiment fertilization affected responses of cuttings to enhanced UV-B radiation, but it concern species and parameters measured. Long-term WUE in P. cathayana and chlorophyll in P. kangdingensis were significantly increased by enhanced UV-B radiation under non-fertilization treatments while the increase was not found under fertilization treatment. In contrast, under no fertilization treatment enhanced UV-B radiation did not affected GPX and APX activity in P. cathayana and GPX in P. kangdingensis while significant increase appeared after application of fertilization. Similar effect of enhanced UV-B radiation on intercellular CO2 concentration in P. kangdingensis was observed.

青杨(Populus cathayana Rehd.)对逆境胁迫的反应及lea基因的表达分析

杨树具有分布广、适应性强，在生态环境治理和解决木材短缺方面均占有重要位置。青杨(Populus cathayana Rehd.)是青杨派树种的重要成员之一，也是我国的特有种。本研究通过对不同水分梯度的干旱胁迫下青杨形态和生理生化的反应，不同pH值盐碱胁迫下不同海拔和不同气候地区的四个青杨种群在生理生态上的反应差异，以及在干旱和低温胁迫下青杨lea2, lea3组基因表达差异的研究，从形态、生理、生化和分子生物学水平系统地研究了青杨在不同逆境胁迫下的反应和青杨不同种群在盐碱胁迫下的反应差异。主要研究结果如下： 1. 青杨在干旱胁迫下的反应机制：中度和重度干旱胁迫下植株的生长受到明显抑制。表现在光合系统上青杨的净光合同化速率(A)下降，主要原因是气孔导度(gs)，胞间二氧化碳浓度(Ci)下降。另外最大量子产量(Fv/Fm)、光化学猝灭效率(qP)降低反应了干旱胁迫下光合系统II(PSII)受到严重损伤, 而且非光化学猝灭效率(qN)上升，导致可利用化学能产量下降，叶绿体产生淀粉的量减少。qP降低qN上升导致产生的过量电子对光合系统的伤害造成活性氧以及丙二醛(MDA)的含量增加。超微解剖结构显示，干旱胁迫增强时，叶绿体内淀粉粒的数目减少，而且叶绿体、线粒体等细胞器中嗜锇颗粒的数目增加。为清除细胞内的活性氧，植物一般的反应是抗氧化系统酶活性增加，对青杨来讲超氧化物歧化酶(SOD), 抗坏血酸过氧化物酶(APx)活性的增加远大于过氧化物酶(POD)，这显示了在青杨中SOD、APx酶在清除活性氧的作用上大于POD。另外同工酶研究结果显示这些酶活性的升高主要是由于各条同工酶带表达量的增加，而不是诱导新酶带的产生。另外，75% FC水分处理下有些指标非但没有下降，像A和有效光量子产量(Y)的值都略有增加，而且gs同时增加。另外，100% FC比75% FC细胞内淀粉粒的数目少一些，但有少量的嗜锇颗粒。这证明100% FC土壤水分也许并非最适合青杨生长。 2. 盐碱胁迫对不同海拔地区青杨种群的反应差异：青杨高海拔和低海拔种群的各种生理特性随着pH值上升都受到了很大的影响。两种群叶和根中Na+、K+ 含量, Na+/K+比率随着pH值的上升影响显著。在pH值高于10.4时高海拔种群叶和根中Na+/K+比率急剧下降但是低海拔种群中却一直维持在较高水平。两种群中MDA、脯氨酸(Proline)的含量，抗氧化系统酶的活性都受到了严重的影响，证明两个种群都属于盐碱胁迫敏感类型但是高海拔的种群对盐碱胁迫的耐性要高于低海拔。这主要是由于高海拔种群一般具有耐干旱、低温胁迫的能力，而植物的抗逆机制一般都有共通之处。 3. 盐碱胁迫对不同气候地区青杨种群的反应差异：盐碱胁迫下两种群的光合作用受到明显的抑制，具体表现在叶绿素的含量和A 显著下降。净光合速率的下降主要是由于叶片gs，Ci 值降低引起的。与湿润地区的种群相比盐碱胁迫增强时，干旱地区的种群叶绿素含量和光合能力的升高与K+离子含量增加有关。植物维持细胞质高K+/Na+值对植物的抗盐性有很重要的作用。为清除盐碱胁迫产生的活性氧，抗氧化系统酶活性增加。盐碱胁迫下干旱地区的种群在SOD、CAT 和谷胱甘肽还原酶(GR)等酶的活性均显著上升，而湿润地区种群只有谷胱甘肽氧化酶(GST)的活性明显增加，说明干旱种群的抗氧化酶系统在较高盐碱胁迫下的保护作用要强于湿润种群。这主要是由于植物抗盐碱胁迫与抗干旱胁迫在一些方面的机制是一致的，抗旱种群一般也能抵抗一定程度的盐碱胁迫。 4. 青杨lea2、lea3 基因在干旱和低温胁迫下的表达差异：通过荧光定量PCR 分析，lea2、lea3 组基因在干旱和低温胁迫下在mRNA 水平的瞬时表达量明显升高，说明了两基因在青杨耐干旱和低温胁迫上都起显著的作用。而且两基因在干旱胁迫下，表达量的升高和降低的时间近乎同步，表明两基因在干旱胁迫下对植物应急保护机制的启动都发挥着重要的作用。低温胁迫下lea3 基因在mRNA 水平上表达量显著上升的时间要早于lea2，而且lea3 基因的持续作用时间明显长于lea2 组基因，说明了低温胁迫开始时lea3基因在植物应对逆境的作用上要大于lea2 基因。 Poplars play an important role in lumber supply, and are important components of ecosystems due to their wide distribution and well adaptation. Populus cathayana Rehd., which belongs to Populus Sect. Tacamahaca Spach, is one of the most important resources of poplars and is specialist to china. In this study, different altitudes and climates populations of P. cathayana were used as experiment materials to investigate the adaptability to drought and salt-alkali stresses. And the cultures of P. cathayana were used to analyze the lea2 and 3 group genes expression when exposed to drought and low temperature stresses. The results are as follows: 1. A large set of parallel responses to drought stress: Drought stress caused pronounced growth inhibition. A decreased significantly and was mainly the result of gs and Ci down. Besides, Fv/Fm, qP decreased and that reflected the harmful effects to PSII of drought stress. In accordance with qN increasing, decreased useful energy production caused the starch numbers reduction in chloroplast. The qP up and qN down improved the levels of ROS and MDA. Starch numbers in chloroplast reduced and plastoglobuli numbers increased when soil water content decreased. To reduce ROS, the activities of SOD, APX, CAT and PPO were activated. The isozymes results show that the rising activities of the antioxidant enzymes resulted from certain isoform content increased, and not from the new band produced. Interestingly, morphological results show 100%FC maybe wasn’t the favorite water content for P. cathayana growth. 2. Effect of salt-alkali stress on morphological and physiological changes in two different altitudes populations of P. cathayana: We compared the physiological responses of two populations of Populus cathayana Rehder, originating from altitudes 2,840 m and 1,450 m. Our results demonstated that Na+ and K+ contents, and Na+/K+ ratios in leaves and roots are greatly affected by pH values. At pH 10.4, the Na+/K+ ratios in both leaves and roots sharply dropped in the higher altitude population but were always maintained at higher levels in the lower altitude population. The pH values causing maximum malondialdehyde (MDA) level, free proline content and antioxidant enzyme activities were significantly different in two populations. These results indicated that the higher altitude population exhibits greater tolerance to alkalinity stress than does the lower altitude population. 3. Morphological and physiological changes in two different climates populations of P. cathayana when exposed to salt-alkali stress. Salt-alkali stress caused pronounced inhibition of the growth and especially in photosystem. Pigments content and A decreased significantly and at the same time gs and Ci decreased too. Compared with wet climate population, the Chlorophyll content and A increased in drought climate population as pH value rising was related to the K+ content increasing. It is important to resist salt-alkali stress that the K+/Na+ ratio matained at high level in cytoplasm. To reduce ROS content, the SOD, CAT and GR activities rised significantly in drought population but only GST increased in wet population. The drought population showed higher salt-alkali tolerance than the wet population mainly resulted from the fact that drought tolerance was in accordance with salt-alkali tolerance to some extent. 4. The different expressional model of lea2 and lea3 gene when P. cathayana was exposed to drought and cold stress. RT-PCR results show both lea2 and lea3 suddenly expressed significantly in mRNA level under drought and cold stress. The expression level of two genes reached optimal level at the same time. But under cold stress, the earlier significantly rising expressional time and the longer maintained higher level time in lea3 than lea2 elucidated that lea3 may be more important than lea2 in resisting cold stress in short time in P. cathayana.

鞑靼荞麦（Fagopyrum tataricum Gaertn.）离体再生与遗传转化研究

鞑靼荞麦是我国特有的农业产品，具有抗寒耐旱特性和较高的营养保健功能。荞麦的开花习性及遗传特点导致其人工杂交授粉难以成功，这成为荞麦杂交育种难以获得突破的重要原因。因此利用转基因技术导入有益基因有可能成为荞麦遗传改良的新途径，而再生及转化体系的建立是开展转基因研究的基础。 本文研究了苗龄、外植体、几种激素配比对鞑靼荞麦（Fagopyrum tataricum Gaertn.）离体培养的影响，初步建立了鞑靼荞麦离体再生体系。结果表明，鞑靼荞麦离体再生的最佳取材时间为苗龄6-8d；诱导愈伤组织的最适培养基为MS＋2.0 mg/L 2,4-D＋1.5 mg/L 6-BA，子叶诱愈率达75％左右，下胚轴的可高达86.62％；愈伤组织分化的最适培养基为MS 0.1mg/L IAA＋2.0mg/L 6-BA＋1.0 mg/L KT＋0.5mg/L TDZ，下胚轴的分化率可达9.52%。下胚轴的诱愈率与分化率均高于子叶，更适于离体再生培养。培养基中加入AgNO3后，能有效降低褐化率。生根最适培养基为含有0.5mg/L NAA的1/2MS培养基，生根率在50％左右。TDZ在诱导鞑靼荞麦的愈伤组织分化出芽的过程中起到明显的促进作用，可提高分化率约20%。 在上述研究基础上，本文还对鞑靼荞麦的遗传转化体系进行了探索性研究。分别利用根癌农杆菌（Agrobacterium tumefaciens）介导法和微粒轰击法（基因枪法）对黑水苦荞下胚轴进行遗传转化。 在农杆菌介导的方法中，携带有质粒pCAMBIA2301的农杆菌菌株EHA105用于转化。载体质粒pCAMBIA2301包含有gus和npt-II 基因, 并受35s启动子驱动。研究结果表明，在侵染方式选择上，浸泡方式比吸打方式更有效，根癌农杆菌侵染的较适浓度为OD600＝0.5，共培养3天，恢复培养7天，能检测到gus基因的表达。 基因枪法使用质粒pBI121，同样包含有gus和npt-II基因, 并受CaMV35s 启动子驱动。轰击距离为9cm较合适，甘露醇前处理在本研究中未表现出明显优势。 两种转化方法比较，基因枪法比农杆菌介导法更快速有效。 本研究为进一步的遗传操作研究打下基础。 Tartary buckwheat (Fagopyrum tataricum Gaertn.), the traditional and unique agricultural product of China, is a kind of crop with strong drought and cold tolerance, abundant nutrition and high medical value. Artificial hybridization is hard in buckwheat because of its flowering habits and genetic characteristics, which leads to no breakthrough in tartary buckwheat breeding. However, biotechnological approaches, especially genetic transformation for the direct introduction of good genes into tartary buckwheat for quality improvement, hold great promise. In this study, we established tartary buckwheat regeneration system in vitro. It is the foundation for genetic manipulation of this crop. The effects of seedling age, hypocotyl and cotyledon as explants, and proportions of several growth regulators were tested in tissue culture of tartary buckwheat for establishing its in vitro regeneration system. The results showed that the best seedling age for callus induction was 6 to 8 days. On the MS medium containing 2.0mg/L 2, 4-D and 1.5mg/L 6-BA, the induction rate of callus from hypocotyls was up to 86.62%, while from cotyledons was about 75％. The suitable shooting medium was the MS medium+0.1mg/L IAA+2.0mg/L 6-BA+1.0 mg/L KT+0.5mg/L TDZ, and the shooting rate from hypocotyls was 9.52%. The callus induction and shooting rates were higher from hypocotyls than from cotyledons. Browning reduced when the medium mixed with AgNO3. Half strength MS supplemented with 0.5mg/L NAA was the best for rooting, the rate was around 50% after 30 days culture. TDZ can accelerate the shoot differentiation distinctively, and it could improve the shooting rate nearly 20%. On the base of above, the explorative research of the genetic transformation in tartary buckwheat was done. In the study, hypocotyls from Heishui tartary buckwheat were transformed by Agrobacterium-mediated method and microprojectile bombardment method (gene-gun), comparatively. In Agrobacterium-mediated method, a disarmed Agrobacterium tumefaciens strain EHA105 harboring plasmid pCAMBIA2301 was used. The vector pCAMBIA2301 contains gus and npt-II genes, driven by CaMV35s promoter. The results showed that the appropriate concentration of Agrobacterium tumefaciens for infecting was OD600＝0.5, and co-culture time was 3d. Seven days later after coculture, GUS expression could be tested. In particle bombardment transformation, plasmid pBI121 was used. pBI121 also contains gus and npt-II genes, driven by 35s promoter. Hypocotyls pretreated with mannitol, no effect was observed, and the suitable distance of bombardment is 9cm. Comparing with Agrobacterium-mediated method, gene-gun method is more convenient and effective. All above results could be a basic work for further study in tartary buckwheat transformation.