超声波辅助提取黑果腺肋花楸花色苷工艺优化及稳定性研究

以黑果腺肋花楸为原料,在单因素实验的基础上应用响应面法对其提取条件进行优化,并研究稳定性。优化黑果腺肋花楸花色苷提取条件为乙醇浓度为63%、超声时间为32 min、超声温度为42℃、料液比为1∶20(g/m L),在此条件下的黑果腺肋花楸花色苷的提取量为5.61 mg/g。在50℃以下的条件下黑果腺肋花楸花色苷稳定性较好,保存率可达98%以上;氧化剂对黑果腺肋花楸花色苷有显著破坏作用;在避光的条件下黑果腺肋花楸花色苷稳定性最好;Ca~(2+)、K~+、Mg~(2+)离子对黑果腺肋花楸花色苷几乎无破坏作用,但Fe~(2+)、Cu~(2+)离子的破坏作用较为显著;在偏酸性条件下黑果腺肋花楸花色苷稳定性较好。     

促生小菇属菌株M23对黑果腺肋花楸光合作用的影响

以黑果腺肋花楸为试验材料,设置5个处理组,分别为阴性对照(CK)、阳性对照(PCK)、大田条件下根施低剂量T₁(每株50 g)、中剂量T₂(每株100 g)和高剂量T₃(每株200 g)3个水平的小菇属菌株M23,分析各处理组黑果腺肋花楸的农艺性状、叶绿素含量和生理指标,并利用Li-6400便携式光合仪测定光合特性日变化。结果表明: 黑果腺肋花楸净光合速率(P_n)呈双峰曲线,在13:00时叶片的气孔导度(g_s)和蒸腾速率(T_r)均明显下降,而胞间CO₂浓度(C_i)却显著升高,出现由非气孔限制因素引起的光合“午休”现象。加菌处理可成功避免光合“午休”现象,与13:00时对照组黑果腺肋花楸的平均值相比,加菌组的平均P_n、g_s、T_r、水分利用率(WUE)和光能利用率(LUE)提高了113%、91%、50%、48%和117%,且日均P_n、g_s、T_r和LUE显著高于对照组,分别约是对照组平均值的1.5、1.9、1.4和1.5倍。在加菌处理组中,高剂量的作用效果显著优于中、低剂量,株高是中、低剂量组的1.2倍。高剂量组黑果腺肋花楸的所有生长指标、光合参数和抗性指标均优于其他组。表明菌株M23可以通过提高黑果腺肋花楸的光合特性、增强对外界环境的适应能力等促进植株生长,且以每株200 g作用效果最佳。     

低pH培养基对发根的促进作用

尽管培养基pH不是关键的控制因素,但一般认为pH5～6是各阶段植物组培物的最适酸度.少数嗜酸植物如乌饭树和杜鹃花则培养在pH4.5～4.8条件下. 匈牙利Fert??d果树及观赏植物研究所的J.M.Zatyko和I.Molnar近期报道,培养在pH3条件下时,黑果腺肋花楸芽培养物产生的根最多.根数量的提高反过来又使芽伸长.除黑果腺肋花楸外,将培养基的pH值调到3～4,还能促进葡萄、木莓、黑莓-木莓和山梣组培物发根.     

黑果腺肋花楸花色苷对人皮肤成纤维细胞紫外辐射损伤的保护作用

为了探讨黑果腺肋花楸花色苷对紫外辐射所致人皮肤成纤维细胞氧化损伤的保护作用,将体外培养的人皮肤成纤维细胞分为对照组、辐射组和辐射给药组。采用MTT法检测不同浓度花色苷添加量对细胞增殖的保护作用,以选择最优添加浓度。采用化学荧光法检测细胞活性氧(ROS)含量,ELISA法检测细胞MMP-1分泌水平。结果表明:UVA辐照剂量为10 J/cm2条件下,与辐射组相比,MTT法显示花色苷添加组浓度为125μg/m L对损伤细胞增殖保护作用有极显著的提高(P0.01),同时,125μg/m L花色苷添加组能够显著降低辐射损伤后细胞ROS含量以及MMP-1分泌水平(P0.01)。由此可见,适宜浓度的黑果腺肋花楸花色苷能够降低细胞ROS含量及MMP-1分泌水平,从而对辐射损的细胞起到一定的保护作用。     

TTC染色鉴定葡萄抗寒性方法的优化与应用北大核心CSCD

以‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’等葡萄砧木和品种直径为0.5~0.7 cm的一年生枝条为材料,经低温梯度(0℃、-14℃、-19℃、-24℃、-29℃和-34℃)处理后,对TTC染色的温度和时间进行优化并观察统计葡萄枝条不同组织的存活情况,测定枝条的相对电导率,以及韧皮部和木质部的可溶性蛋白、游离脯氨酸、可溶性糖、淀粉含量和束缚水自由水比例(束自比)5个生理指标,分别用枝条纵切面染色面积、相对电导率拟合Logistic方程计算枝条的半致死温度(LT_(50))来评价枝条的抗寒性,同时通过隶属函数法综合评价枝条韧皮部和木质部抗寒性,并将3种方法的评价结果进行比较,以建立一种直观高效鉴定葡萄品种抗寒性的方法。结果表明:(1)依据低温胁迫下各品种葡萄砧木扦插枝条的萌芽率和生根率表现,其抗寒性由强到弱依次为‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’。(2)葡萄枝条TTC活力染色的最佳条件为pH=7.0,0.5%TTC-0.1 mol/L磷酸缓冲液在35℃下避光染色36 h。随着胁迫温度的降低,各品种枝条纵切面染色面积均逐渐降低,根据优化TTC法获得‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’枝条的LT_(50)分别为-31.38℃、-26.51℃、-26.10℃、-23.60℃、-23.33℃和-19.26℃。(3)随着胁迫温度的降低,各品种枝条的相对电导率逐渐增加,且‘黑比诺’的相对电导率基本保持最高、增幅最大(51.93%),而‘左山1号’的相对电导率始终最低、增幅最小(44.07%);根据相对电导率获得‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’枝条的LT_(50)分别为-30.02℃、-26.40℃、-25.75℃、-23.16℃、-21.13℃和-17.72℃。(4)通过5种生理指标进行枝条抗寒性隶属函数综合性评价结果显示,同一品种中韧皮部抗寒性强于木质部,同一部位不同品种的抗寒性表现为:‘左山1号’>101-14>3309C>SO4>140R>‘黑比诺’。研究发现,TTC染色法与电导法和综合隶属函数法获得的葡萄枝条抗寒力评价结果一致,也与枝条生长恢复鉴定结果一致;但与其他两种方法相比,TTC染色法能更加直观和有效地预测评估葡萄枝条的抗寒性。     

黑果腺肋花楸的离体快繁技术

1　植物名称　黑果腺肋花楸 (Aroniamelanocarpa)。2　材料类别　顶芽、侧芽。3　培养条件　 ( 1 )芽诱导培养基 :MS + 6 BA 0 .5mg·L- 1 (单位下同 ) +NAA 0 .2 ;( 2 )增殖继代培养基 :MS + 6 BA 1 .0 +NAA 0 .0 8;( 3)生根培养基 :1 /2MS+ 6 BA 0 .5 +IBA 0 .3。上述培养基均含3.5 %食用白糖、0 .8%食用琼脂 ,pH 5 .9。培养温度为 ( 2 5± 2 )℃ ,光照度 2 5 0 0～ 32 0 0lx ,光照时间1 6h·d- 1 。4　生长与分化情况4.1　无菌材料的获得　春季 3月下旬～ 4月中旬 ,取二年生枝条 ,先在洗衣粉水中用毛刷刷洗 ,再用流动水冲洗 1 …     

黑果腺肋花楸花色苷通过Nrf2机制对Aβ_(1-42)诱导SH-SY5Y细胞产生氧化应激及细胞凋亡的保护作用

为了研究黑果腺肋花楸花色苷对Aβ_(1-42)诱导的SH-SY5Y细胞产生氧化应激和细胞凋亡的影响。采用MTT比色法测定MTT,分别用ROS、H_2O_2、SOD检测试剂盒测定活性氧(ROS),过氧化氢(H_2O_2),超氧化物歧化酶(SOD)等氧化应激相关指标。采用Annexin-V-PI/FITC凋亡检测试剂盒检测细胞凋亡。采用RT-PCR,蛋白质印迹法分析抗氧化酶相关基因核转录相关因子(Nrf2)、血红素氧化酶-1(HO-1)、醌氧化还原酶(NQO1)、凋亡相关基因(Bcl-2、Bax)的转录和蛋白表达。结果表明花色苷预处理显著抑制Aβ_(1-42)诱导的细胞内ROS、H_2O_2的增加,并通过上调Nrf2、HO-1、NQO-1表达来增加SOD表达。通过下调Bax,上调Bcl-2显著抑制细胞凋亡。从而得出高纯度的黑果腺肋花楸花色苷可以通过Nrf2机制保护SH-SY5Y细胞免受Aβ_(1-42)诱导的氧化应激和细胞凋亡。     

腊梅的环剥扦插繁殖法

腊梅的环剥扦插繁殖法腊梅（ＣｈｉｍｏｎａｎｔｈｕｓｐｒａｅｃｏｘＬｉｎｋ）属腊梅科,落叶灌木。单叶对生,卵状椭圆形,表面粗糙。秋季孕蕾,花由一年生枝条叶腋发生,先花后叶,花期１２月至次年１～２月;花单生,两性、黄色带蜡质,果椭圆形。腊梅原产我国中部各...     

黑果腺肋花楸的组织培养和快速繁殖

1植物名称黑果腺肋花楸(Aroniamelanocarpa). 2材料类别茎尖、茎段. 3培养条件(1)初始诱导培养基:1/3MS 6-BA 1.0 mg·-1(单位下同) IBA 0.2;(2)不定芽诱导培养基:MS 6-BA 1.0 IBA 0.2;(3)增殖培养基:MS IBA 0.2～0.5 IBA 0.2;(4)生根培养基:1/2MS IBA 0.5或者1/2MS NAA0.5.上述培养基中均加入2%～3%的蔗糖和0.6%的琼脂粉,pH 5.8.培养温度20～25℃,光照度1 500～2 000 lx,光照时间12 h·d-1.     

银杏嫁接早结实

银杏是著名的经济树种。过去银杏生产一直采取实生繁育,不选种、不嫁接,导致银杏个体良萎不齐、结实晚、品质差、产量低。为了改良品种,早实丰产,我们从１９８９年起,进行了银杏不同嫁接方法和嫁接时期的试验研究。总结出成活率高、嫁接期长、操作简便的三种嫁接方法。１．劈接具有延长嫁接期、生长势强等特点。可分为春接和秋接。春接宜在３月上旬至４月上旬进行,秋接直在吕月中旬至９月中旬进行。对接穗的要求（三种方法都类似）,春接于２月下旬至３月下旬,采集优良品种母树上枝条,如嵩优２号、嵩优４号、嵩优１８号、中银黑１号…     

Extended alternating-temperature cold acclimation and culture duration improve pear shoot cryopreservation

Meristems of many pear genotypes can be successfully cryopreserved following 1 week of cold acclimation, but an equal number do not survive the process or have very little regrowth. This study compared commonly used cold acclimation protocols to determine whether the cold acclimation technique used affected the cold hardiness of shoots or the regrowth of cryopreserved meristems. In vitro-grown pear (Pyrus L.) shoots were cold acclimated for up to 16 weeks, then either the shoot tips were tested for cold hardiness or the meristems were cryopreserved by controlled freezing. Cold acclimation consisted of alternating temperatures (22 degrees C with light/-1 degrees C darkness with various photo- and thermoperiods) or a constant temperature (4 degrees C with an 8-h photoperiod or darkness). Compared with nonacclimated controls, both alternating- and constant-temperature acclimation significantly improved postcryopreservation regrowth of P. cordata Desv. and P. pashia Buch. -Ham. ex D. Don meristems. Alternating-temperature acclimation combined with either an 8-h photoperiod or darkness was significantly better than constant-temperature acclimation. Alternating-temperature shoot acclimation for 2 to 5 weeks significantly increased postcryopreservation meristem regrowth, and recovery remained high for up to 15 weeks acclimation. Postcryopreservation meristem regrowth increased with 1 to 5 weeks of constant-temperature acclimation and then declined with longer acclimation. Shoot cold hardiness varied with the acclimation procedure. The LT(50) of shoots acclimated for 10 weeks with alternating temperatures was -25 degrees C; that with constant temperature was -14.7 degrees C; and that of the nonacclimated control was -10 degrees C. Less frequent transfer of cultures also improved acclimation of shoots. Shoots grown without transfer to fresh medium for 6-12 weeks had higher postcryopreservation recovery with shorter periods of acclimation than shoots with a 3-week transfer cycle.     

Effect of photoperiod and temperature on the development of frost hardiness in three Alnus species

The influence of short day and low temperature on cold acclimation of A. crispa (Ait.) Pursh, A. glutinosa (L.) Gaertn. and A. rubra Bong, was investigated. Two clones of each species originating from in vitro propagation were exposed to three daylength/temperature treatments. Periodically plantlets were exposed to controlled freezing temperature in order to evaluate their level of frost hardiness.
Short day (SD) and cold temperature (CT) and long day (LD) and cold temperature (CT) were the most effective treatments for the development of frost hardiness in shoots and roots of the three species tested. Short day (SD) and warm temperature (WT) induced a significant increase in hardiness in shoots of all three species. However, this treatment did not trigger root hardening. A. crispa was found to be the hardiest species followed by A. glutinosa and A. rubra . Intraspecific variation was observed between the two A. glutinosa clones. A glutinosa clone AG8, a Russian provenance, showed a greater freezing resistance than A. glutinosa clone AG2, a German provenance.     

Cold hardening of raspberry plants in vitro is enhanced by increasing sucrose in the culture medium

The influence of exogenously applied sucrose on cold hardening of raspberry ( Rubus idaeus L.) in vitro was examined. Raspberry plants (cv. Preussen) were cultured on Murashige-Skoog (MS) media with different levels (1, 3, 5 and 7%) of sucrose and subjected to low-temperature acclimation (3/−3°C day/night temperature, 8-h photoperiod) for 14 days. Cold hardiness (LT₅₀ in controlled freezing), shoot moisture content, osmolality and the amounts of sucrose, glucose and fructose were determined. Exogenously applied sucrose was taken up by plants, but the uptake corresponded to less than 10% of total sugar reserves in the culture. Cold hardiness was primarily affected by acclimation treatment, but sucrose increased cold hardiness of nonacclimated plants and significantly enhanced the effect of acclimation treatment, 5% sucrose in the culture medium being optimal for cold hardening. LT₅₀ values ranged between −4.1 and −7.1°C for nonacclimated, and between −14.2 and −20.7°C for cold-acclimated shoots. Shoot moisture content was inversely related to medium sucrose level and declined only slightly during cold acclimation. After cold acclimation, plant osmolality predicted hardiness better than shoot moisture content. Plant osmolality and sugar content were increased by increasing the medium sucrose level and, to a greater extent, by cold acclimation. Sucrose, glucose and fructose accumulated during hardening. Sucrose was the predominant sugar, and the rate of sucrose accumulation during cold acclimation was independent of the medium sucrose level or the initial plant sucrose content. A close correlation between cold hardiness and total sugars, sucrose, glucose and fructose was established. These results suggest that sugars have more than a purely osmotic effect in protecting acclimated raspberry plants from cold.     

Metabolic dynamics during autumn cold acclimation within and among populations of Sitka spruce (Picea sitchensis)

? Autumnal cold acclimation in conifers is a complex process, the timing and extent of which vary widely along latitudinal gradients for many tree species and reflect local adaptation to climate. Although previous studies have detailed some aspects of the metabolic remodelling that accompanies cold acclimation in conifers, little is known about global metabolic dynamics, or how these changes vary among phenotypically divergent populations. ? Using untargeted GC-MS metabolite profiling, we monitored metabolic dynamics during autumnal cold acclimation in three populations of Sitka spruce from the southern, central, and northern portions of the species range, which differ in both the timing and extent of cold acclimation. ? Latitudinal variation was evident in the nature, intensity, and timing of metabolic events. Early development of strong freezing tolerance in the northern population was associated with a transient accumulation of amino acids. By late autumn, metabolic profiles were highly similar between the northern and central populations, whereas profiles for the southern population were relatively distinct. ? Our results provide insight into the metabolic architecture of latitudinal adaptive variation in autumn acclimation and show that different mechanisms are the basis of early October cold hardiness and autumn-acclimated cold hardiness.     

Deep Undercooling in Woody Taxa Growing North of the -40 degrees C Isotherm

The freezing of deep undercooled water in cold-hardened 3-year-old stems of 16 woody taxa was studied in mid-January by differential thermal analysis. The initiation temperature and the size of the low temperature exotherm (LTE) were compared for nonthawed, thawed, and freeze-killed stems. In general, the initiation temperature of the LTE for nonthawed stems occurred at a lower temperature than for thawed stems and freeze-killed stems. In some cases, no LTE was detected in nonthawed stems although a LTE was detected after thawing. The size of the LTE increased after thawing the stem and also after the stem was freeze killed. The LTE observed in one species disappeared upon exposure to continuous low sub-zero temperatures. Results suggest that undercooling which subsequently results in the LTE in woody stems is due to the cell wall and the plasma membrane. During periods of prolonged freezing, cellular water migrates from the cells which undercool to extracellular ice. This results in a concentration of cell solutes which lowers the homogeneous nucleation temperature of the cell sap. The cold hardiness of nonthawed and thawed stems was compared by a controlled freeze test. In general, thawing had little effect on the survival temperature whereas it had a marked effect on the initiation of the LTE.     

Low night temperature and inhibition of gibberellin biosynthesis override phytochrome action and induce bud set and cold acclimation, but not dormancy in PHYA overexpressors and wild-type of hybrid aspen

Juvenile trees of temperate and boreal regions cease growth and set buds in autumn in response to short day-lengths (SD) detected by phytochrome. Growth cessation and bud set are prerequisites for the development of winter dormancy and full cold hardiness. In this study we show that the SD-requirement for bud set and cold hardening can be overcome in hybrid aspen (Populus tremula L. × tremuloides Michx.) by low night temperature and inhibition of gibberellin (GA) biosynthesis. Bud set and increased cold hardiness were observed under normally non-inductive long day-length (LD) in wild-type plants, when exposed to low night temperature and paclobutrazol. In addition, the effect of PHYA overexpression could be overcome in transgenic plants, producing bud set and cold acclimation by treatment with: SD, low night temperature and paclobutrazol. After cold acclimation, the degree of bud dormancy was lower for wild-type plants prior treated with LD and transgenic plants (overexpressing PHYA), than SD-treated, wild-type plants. Thus, low night temperature in combination with reduced GA content induced bud set and promoted cold hardiness under normally non-inductive photoperiods in hybrid aspen, but was unable to affect development of dormancy. This might suggest separate signalling pathways from phytochrome regulating the induction of cold/cold hardiness and bud dormancy in hybrid aspen or alternatively, there might be one pathway that fails to complete its action in the transgenic and paclobutrazol treated plants.     

Effect of frost nights and day and night temperature during dormancy induction on frost hardiness, tolerance to cold storage and bud burst in seedlings of Norway spruce

For trees, the ability to obtain and maintain sufficient levels of frost hardiness in late autumn, winter and spring is crucial. We report that temperatures during dormancy induction influence bud set, frost hardiness, tolerance to cold storage, timing of bud burst and spring frost hardiness in seedlings of Norway spruce (Picea abies (L.) Karst.). Bud set occurred later in 12°C than in 21°C, and later in cool nights (7°C) than in constant temperature. One weekly frost night (−2.5°C) improved frost hardiness. Cool nights reduced frost hardiness early, but improved hardiness later during cold acclimation. Buds and stems were slightly hardier in 21°C than in 12°C, while needles were clearly hardier in 12°C. Cold daytime temperature, cool nights and one weekly frost night improved cold storability (0.7°C). Seedlings receiving high daytime temperatures burst buds later, and were less injured by light frost some days after bud burst.     

Does ice crystal formation in buds explain growth disturbances in boron-deficient Norway spruce?

Loss of apical dominance in boron-deficient trees has been suggested to be due to frost damage of terminal buds and leaders. Excessive nitrogen (N) supply can exacerbate boron (B) deficiency by the dilution-effect. N may also have direct effects on winter hardiness. We studied frost hardening of buds of Norway spruce (Picea abies L. Karst.) in healthy-looking trees and in trees with growth disturbances. The effect of B and N on frost hardiness was studied in a factorial fertilisation experiment during cold acclimation. Frost hardiness was determined by differential temperature analysis (DTA) and scoring of visual damage. In a DTA profile of apical buds with a piece of stem, low-temperature exotherm (LTE) predicted bud injury, while two of the observed high-temperature exotherms and two of the observed intermediate-temperature exotherms were non injurious. Appearance of LTE followed changes in air temperature. The risk of frost damage was not affected by fertilisation treatments or previously observed growth disturbances. However, when the bud structure was deformed by severe B deficiency, the supercooling ability disappeared. Such buds are probably killed by freezing in nature and therefore, frost damage may play a secondary role in the development of growth disturbances.     

The environmental control of cold acclimation in apple

The role of photoperiod and temperature in the cold acclimation of living Haralson apple (Pyrus malus L.) bark was studied in the autumn under field conditions in Minnesota. Whole trees, or different parts of the same tree, were exposed to either natural conditions, artifically lengthened days, or artificially warmed nights, or they were subjected to manual leaf removal. The results indicate that acclimation occurs in two stages which are induced by short days and frost (or low temperature), respectively. Leaves were stimulated by short days to produce translocatable substance(s) which promoted cold acclimation of the living bark. Leaves of plants grown under long days were the source of a translocatable substance(s) which inhibited acclimation. The second stage of hardiness, induced by frost (or low temperature), did not involve translocatable factors.     

Cold Hardiness and Acclimation Intensity in Flower Buds for Fall Bloom and Spring Bloom Clones in Rhododendron kiusianum, a Dwarf Evergreen Azalea

The relationship between the degree of cold hardiness (supercoolingability of florets) and the acclimation intensity in flowerbuds was investigated in the fall bloom and the spring bloom(typical) clones of Rhododendron kiusianum, a hardy dwarf evergreenazalea. Supercooling ability or exotherm temperature distribution(ETD) in florets was determined by differential thermal analysis(DTA) and the intensity of bud acclimation or the rate of deacclimationwas judged by the changes in ETD profiles resulting from thedehardening temperature treatment. Although the two clone typesshowed no significant differences in ETDs and water contentsin florets, they differed in their rates of bud deacclimation.The flower buds of fall bloom clones generally tend to deacclimatemore quickly than the spring bloom ones throughout the seasons.It is concluded that the degree of cold hardiness in flowerbuds of R. kiusianum does not differ between the fall bloomand spring bloom clones but the intensity of bud acclimationdoes; acclimation intensity is higher in the spring bloom clonesand the rate of deacclimation is greater in the fall bloom ones. (Received October 14, 1985; Accepted February 5, 1986)