梅花(Prunus mume Sieb.et Zucc.)远缘杂种及其亲本的差热分析与冻害关系的研究

用差热分析(Differential thermal analysis: DTA)研究了山桃(Prunus davidiana)、杏(P. armeniaca)、青岛“粉红梅”(P.mume cv.' Fenhong Mei,)、“小绿萼”(P.mume cv.'Small Green Calyx')及其种问杂种“小绿萼”梅×山桃、青岛“粉红梅”×杏和杏×青岛“粉红梅”的低温放热(Low temperature exotherm)与冻害关系,以及皮部和木质部的冰冻类型(Freezing pattern)。在差热分析中,观察到亲本和杂种的木质部都有二次放热现象。低温放热后,引起木质部和髓射线薄壁细胞死亡,原生质膜透性急剧增加。在杂种与亲本之间,存在着明显的差异。分离的皮部却只出现一次高温放热(High temperature exotherm)。高温放热是与冻害无关的。文末讨论了梅花及其杂种在北京越冬的主要障碍及有关栽培措施。     

梅与山桃远缘杂交亲和性初步研究

为培育早花抗寒梅花新品种,以梅(Prunus mume)品种‘江梅’(P.mume‘Jiangmei’)、‘淡丰后’(P.mume‘Dan Fenghou’)与山桃(P.davidiana)、‘白花’山桃(P.davidiana‘Alba’)为亲本进行杂交试验,记录种间杂交结实率,观察花粉管生长,对未成熟胚进行培养。结果表明:(1)梅与山桃、‘白花’山桃杂交结实率很低,‘江梅’ב白花’山桃未结果,结实率最高的组合为‘淡丰后’×山桃,也仅有7.4%,且杂交果实的果核内部分胚干瘪、败育。(2)山桃和‘白花’山桃的花粉在梅柱头上都能正常萌发,但花粉管生长受抑制,多数花粉管到达花柱中部即弯曲、缠绕、断裂,花粉管生长过程中有大量的胼胝质产生,表现较低的杂交亲和性,但不同种间杂交亲和程度又有所不同。(3)通过未成熟胚培养获得了杂种苗。研究表明,梅与同属种杂交存在不亲和性,幼胚拯救是获得梅与李属其他种远缘杂交杂种苗的有效途径。     

Typha×glauca Godr.,香蒲属(香蒲科)中国新记录杂种及其形态特征

香蒲属(Typha L.)为多年生水生或沼生草本植物, 种间存在十分普遍的杂交现象, 其中一些杂种在湿地生态系统中有重要的作用。在查阅大量腊叶标本基础上, 结合野外居群生物学工作, 作者发现中国一新记录杂种, 即T.×glauca Godr. (T. angustifolia L.×T. latifolia L.), 并新拟“粉绿香蒲”作该杂种的中文名。对粉绿香蒲的形态特征进行了研究, 讨论了其与亲本水烛和宽叶香蒲的区别, 并给出检索表。     

基于核基因和叶绿体基因序列的杏属系统发育分析——探讨洪平杏的起源和亲缘关系

洪平杏(Armeniaca hongpingensis C. L. Li)是杏属的一个狭域分布种,基于形态观察被推测为杏(A.vulgaris Lam.)和梅(A. mume Sieb.)的天然杂交种,但目前尚无该种与杏、梅亲缘关系的分子系统学研究。本文以洪平杏的成株和实生苗以及包括杏、梅在内的6种(含1变种)杏属植物为研究材料,分别采用核基因(ITS和SBEI)和叶绿体基因(mat K和ycf1b)序列构建系统发育树,并采用mat K、ycf1b和SBEI基因序列构建单倍型网络图,探讨该物种与杏、梅及杏梅(A. mume Sieb. var. bungo Makino)之间的亲缘关系。基于核基因和叶绿体基因序列分别构建的系统发育树均显示,洪平杏的成株及其全部实生苗个体单独聚为一支,且具有较高的支持率(分别为99/79、71/81),独立于杏属其他种之外。而基于核基因ITS序列的系统发育分析结果表明,洪平杏除极少数成株与杏、杏梅聚为一支外,其余所有成株与实生苗聚为2大支(支持率分别为0.82和0.97),而没有克隆的与梅聚在一起。单倍型分析结果表明,该物种的成株与实生苗在SBEI和ycf1b基因序列中均未检测到杏或梅的单倍型,仅有少数(2/9)的实生苗个体在叶绿体mat K基因序列中检测到杏的单倍型。研究结果不支持将洪平杏定为杏和梅的天然杂交种的观点,推测洪平杏应为一个独立的物种,与杏之间的亲缘关系更近并且存在可检测到的基因流。     

基于30个形态性状的中国杏属（Armeniaca Scop.）植物分类学研究

基于与叶和果实相关的30个形态性状(包括20个定性性状和10个定量性状)对中国杏属( Armeniaca Scop.)11种3变种进行了UPGMA聚类分析和主成分分析;在主成分分析基础上,构建了中国杏属植物的OTU散点图;并且,结合降水量分布图绘制了中国杏属植物分布图。聚类分析结果显示：供试杏属植物被分成2支。若包含毛叶梅( A. mume var. goethartiana Koehne),则毛叶梅、梅( A. mume Sieb.)、洪平杏( A. hongpingensis C. L. Li)以及政和杏( A. zhengheensis J. Y. Zhang et M. N. Lu)聚为一支,其余8种2变种聚为另一支;若不包含毛叶梅,梅则被划分在后一个分支中。主成分分析结果显示：前3个主成分的累计贡献率仅60.3180%,说明中国杏属植物的形态性状具有较大的遗传变异;在前3个主成分中,树高、叶片下表面被毛情况、叶长/叶柄长比值、叶长/叶宽比值、果核形状、叶宽、果核宽、叶柄长、果柄长和叶缘锯齿形状的绝对权重值均在0.7以上,表明这10个性状在中国杏属植物的分类学研究中具有重要作用。 OTU散点图显示：中国杏属植物在二维散点图上的分类结果与其聚类结果基本一致,并且,其聚类结果中的各分支在三维散点图上也能够明显区分,说明可以采用前3个主成分中绝对权重值较高的性状对中国杏属植物进行分类。分布图显示：杏属植物遍布中国各省(区),且主要集中分布在400和800 mm等降水量线之间的区域。结合上述研究结果及他人的研究成果,支持“将藏杏﹝A. holosericea ( Batal.) Kost.〕作为杏( A. vulgaris Lam.)的1个变种”以及“将政和杏作为梅的1个变种”的分类处理,并支持“将洪平杏作为独立种”的分类处理。此外,建议将仙居杏( A. xianjuxing J. Y. Zhang et X. Z. Wu)和华仁杏( A. cathayana D. L. Fu et al)作为杏属的栽培种。     

两个葡萄杂交后代根系抗葡萄根瘤蚜及抗寒性鉴定

【目的】为了筛选抗葡萄根瘤蚜Daktulosphaira vitifoliae Fitch且抗寒的葡萄砧木以适应我国葡萄生产需求。【方法】以山葡萄Vitis amurensis Rupr.左山1号×SO4杂种F1代的45个株系(A系列)和左山1号×101-1杂种F1代27个株系(B系列)为试材,采用离体根接种鉴定法进行抗葡萄根瘤蚜鉴定及抗性分级;采用差热分析系统(differential thermal analysis,DTA)进行各株系根系的低温放热(low temperature exotherms,LTE)分析,建立各株系根系韧皮部及木质部的温度-伤害度(LT-I)回归方程,评估各株系根系的抗寒性。【结果】葡萄根瘤蚜在杂交株系根系上的产卵量均显著低于敏感品种巨峰,筛选出被葡萄根瘤蚜侵染后不能形成根瘤,抗葡萄根瘤蚜级别为0级的A系列杂交株系18个和B系列株系11个。被葡萄根瘤蚜侵染后形成根瘤比例低于10%的抗葡萄根瘤蚜级别为1级的A系列杂交株系9个和B系列株系4个;筛选出A系列综合低温放热温度隶属度函数、韧皮部和木质部低温放热温度隶属度函数3个指标均低于贝达的株系27个,B系列各指标均低于贝达的株系3个。【结论】本研究筛选出抗寒性强且对葡萄根瘤蚜抗性强的A系列株系15个和B系列株系2个。其中,A14,A16,A18,A22,A23,A28,A34,A35,A38,A44,A50,B24和B26对葡萄根瘤蚜抗性级别为0;A11,A15,A17和A27对葡萄根瘤蚜抗性级别为1。     

夏蜡梅和美国蜡梅及属间杂种‘红运’营养器官解剖结构特征比较

采用石蜡切片法对夏蜡梅〔Sinocalycanthus chinensis(Cheng et S.Y.Chang)Cheng et S.Y.Chang〕和美国蜡梅(Calycanthus floridus L.)及属间杂种‘红运’(S.chinensis×C.floridus‘Hong Yun’)叶、茎和根的横切面解剖结构进行了比较观察。观察结果显示:夏蜡梅、美国蜡梅及杂种‘红运’的叶、茎和根的横切面解剖结构有较高的共性,但仍有明显区别。杂种‘红运’的叶片厚度、主脉维管束横截面面积和海绵组织厚度与夏蜡梅比较接近,而与美国蜡梅差异较大;杂种‘红运’叶的栅栏组织厚度与海绵组织厚度的比值(P/S)和栅栏组织厚度与叶片厚度的比值(P/L)均介于2个亲本之间,P/S和P/L 2个指标均为美国蜡梅最高、夏蜡梅最低。杂种‘红运’茎的直径、皮层细胞层数、皮层厚度、皮层厚度与茎半径的比值、木质部厚度和维管束面积等指标的数值均大于美国蜡梅和夏蜡梅。美国蜡梅根的髓部不发达,夏蜡梅根则有明显的髓部,而杂种‘红运’根的髓部较发达;美国蜡梅根的木质部和木栓层均最厚,髓部分布的大型导管数量最多;杂种‘红运’根的皮层最发达,木质部最不发达,髓部的大型导管最少。根据营养器官的解剖结构特征与生态特性的关系推测:夏蜡梅具有不喜强光、稍耐阴的生态特性,美国蜡梅的耐热性和光合能力最强,而杂种‘红运’比其母本夏蜡梅有更好的生态适应性。     

甘蓝型油菜(Eru CMS)与甘蓝种间杂种的同工酶和蛋白质分析

经胚抢救获得甘蓝型油菜(Brassica napus L.)(Eru CMS)与甘蓝(Brassica oleracea L.var.capitata L.)种间杂种,前期经过流式细胞仪、柱头染色体数目、花粉活力等分析获得一些真杂种。利用电泳法,对真杂种植株的3种同工酶(SOD、EST、COD)和蛋白质进行详细分析,了解了杂种与亲本的同工酶和蛋白质的特性差异。结果表明,杂种与亲本之间的同工酶和蛋白质存在较明显的差异:杂种的SOD、COD的酶带表现为偏父本甘蓝型;杂种的EST的酶带表现为偏母本油菜型;杂种的蛋白质电泳表现为不仅具有双亲的特征蛋白带,也有其自身特征蛋白带。     

薯蓣属植物人工杂交后代的检测

以湖北武当山的盾叶薯蓣(Dioscorea zingiberensis C.H.Wright)为主要亲本,与重庆金佛山的盾叶薯蓣、小花盾叶薯蓣(D.parviflora C.T.Ting)及黄独(D.bulbifera L.)进行种内和种间杂交,并获得部分杂交组合的F1代植株。将F0代种子与F1代植株进行结实率及种子萌发率比较,并与亲本进行过氧化物酶(POD)、超氧化物歧化酶(SOD)和酯酶(EST)同工酶比较。结果表明,不同亲本及其杂交后代的结实率、种子萌发率和实生苗存活率均存在明显差异;亲本和F1代的同工酶酶谱较丰富,部分杂种与亲本的相似度较高,并出现各自的特征酶带。运用同工酶技术可以鉴别杂种的真伪。     

干旱胁迫对鸭茅幼苗根系生长及光合特性的影响

以不同耐旱型鸭茅品种(系)“01998”(敏感型)和“宝兴”(耐旱型)为试材,采用盆栽控水试验方法,研究土壤干旱胁迫对鸭茅幼苗根系生长、生理特性及叶片光合作用的影响,探讨不同品种耐旱机制的差异.结果表明：干旱胁迫下,不同耐旱型鸭茅幼苗的根系活力和根系数量均呈先上升后下降的趋势,当土壤相对含水量降到30%时,根系活力和根数显著增加并达到最大值;干旱胁迫降低了两个品种鸭茅叶片的相对含水量、叶绿素含量、净光合速率、叶片蒸腾速率、气孔导度,提高了电导率和胞间CO₂浓度.干旱胁迫降低了两个品种鸭茅的单株叶面积,品种“01998”的地下及地上生物量下降,而“宝兴”无显著变化.     

Cold Acclimation in Genetically Related (Sibling) Deciduous and Evergreen Peach (Prunus persica [L.] Batsch): I. Seasonal Changes in Cold Hardiness and Polypeptides of Bark and Xylem Tissues

Seasonal patterns of proteins and of cold hardiness were characterized in bark and xylem tissues of genetically related (sibling) deciduous and evergreen peach (Prunus persica [L.] Batsch). In contrast with deciduous trees, which entered endodormancy and abscised leaves in the fall, evergreen trees retained their leaves and exhibited shoot elongation under favorable environmental conditions. A successive increase in the cold hardiness of bark and xylem was observed during the fall in both genotypes. This was followed by a subsequent decrease from midwinter to spring. Xylem tissue in both genotypes exhibited deep supercooling and a significant correlation (r = 0.99) between the midpoint of the low-temperature exotherm and the subzero temperature at which 50% injury occurred (assessed by electrolyte leakage) was noted. The maximum hardiness level attained in deciduous trees was more than twofold that of evergreens. Seasonal pattern of proteins from bark and xylem of the sibling genotypes was characterized by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among other qualitative and quantitative changes, accumulation of a 19-kilodalton polypeptide in the bark of both genotypes was observed during fall followed by a decrease in spring. This polypeptide accumulated to higher levels in the deciduous peach compared with the evergreen. Additionally, a 16-kilodalton protein exhibited the same pattern in deciduous trees but not in the evergreen trees. Both the 19- and a 16-kilodalton bark proteins conform to the criteria of a bark storage protein. The relationship of seasonal changes in protein to cold hardiness and dormancy in these genetically related peach genotypes is discussed.     

Freezing Events within Overwintering Buds of Blackcurrant (Ribes nigrum L.)

Overwintering buds of blackcurrant cultivars 'Ben Lomond' and'Ben More' were examined by differential thermal analysis (DTA).Photographic evidence relates the first (primary) exotherm tothe freezing of water in the basal pith and bud scales. Thenumber of secondary exotherms either matched, or was fewer than,the number of floral racemes within the bud. There is evidencein the structure of the secondary exotherms that the freezingof individual primordia was being recorded.Copyright 1993, 1999Academic Press Differential thermal analysis, freezing injury, buds, Ribes nigrum, blackcurrant     

Cold hardiness and deep supercooling in xylem of shagbark hickory

Differential thermal analysis, differential scanning calorimetry, pulsed nuclear magnetic resonance spectroscopy, and low temperature microscopy are utilized to investigate low temperature freezing points or exotherms which occur near −40 C in the xylem of cold-acclimated shagbark hickory (Carya ovata L.). Experiments using these methods demonstrate that the low temperature exotherm results from the freezing of cellular water in a manner predicted for supercooled dilute aqueous solutions. Heat release on freezing, nuclear magnetic resonance relaxation times, and freezing and thawing curves for hickory twigs all point to a supercooled fraction in the xylem at subfreezing temperatures. Calorimetric and low temperature microscopic analyses indicate that freezing occurs intracellularly in the xylem ray parenchyma. The supercooled fraction is found to be extremely stable, even at temperatures only slightly above the homogeneous nucleation temperature for water (−38 C). Xylem water is also observed to be resistant to dehydration when exposed to 80% relative humidity at 20 C. D₂O exchange experiments find that only a weak kinetic barrier to water transport exists in the xylem rays of shagbark hickory.     

The mechanism of freezing injury in xylem of winter apple twigs

In acclimated winter twigs of Haralson apple (Pyrus Malus L.), a lag in temperature during cooling at a constant rate was observed at about −41 C by differential thermal analysis. The temperature at which this low temperature exotherm occurred was essentially unaffected by the cooling rate. During thawing there was no lag in temperature (endotherm) near the temperature at which the low temperature exotherm occurred, but upon subsequent refreezing the exotherm reappeared at a somewhat higher temperature when twigs were rewarmed to at least −5 C before refreezing. These observations indicate that a small fraction of water may remain unfrozen to as low as −42 C after freezing of the bulk water in stems. The low temperature exotherm was not present in twigs freeze-dried to a water content below 8.5% (per unit fresh weight), but it reappeared when twigs were rehydrated to 20% water. When freeze-dried twigs were ground to a fine powder prior to rehydration, no exotherm was observed. Previous work has shown that the low temperature exotherm arises from xylem and pith tissues, and that injury to living cells in these tissues invariably occurs only when twigs are cooled below, but not above the temperature of the low temperature exotherm. This study revealed that the low temperature exotherm resulted from the freezing of a water fraction, that the freezing of this water was independent of the freezing of the bulk water, that the exotherm was associated with some gross structural feature but not the viability of the tissue, and that injury to living cells in the xylem and pith was closely and perhaps causally related to the initial freezing of this water.     

Comparative freezing patterns of bark and xylem of “Siberian C” and “Redhaven” peach twigs

An electrophoretic mobility technique was used to study freezing patterns in excised peach twigs. Moisture content was the only qualitative difference in initial freezing patterns of similar tissues of Redhaven and Siberian C. Siberian C contained up to 18% less moisture than Redhaven. Major differences in the shape of the transition pattern were detected between bark and xylem. Even though bark tissues had twice the water content of xylem, the bark exhibited equilibrium freezing while the xylem underwent nonequilibrium freezing. Bark water must be intimately associated with the living protoplast, while xylem water is less closely associated with cellular components. Comparison of bark and xylem freezing curves with sucrose and cellulose model systems suggested that bark freezing was similar to the sucrose model while xylem freezing was similar to the cellulose model.     

Low temperature exotherms of winter buds of hardy conifers

Differential thermal analysis (DTA) of winter buds and the excisedprimordial shoots of sub-alpine or sub-cold firs revealed thatthese buds had all low temperature exotherms around –30?C.However, no low temperature exotherm below –15?C was detectedin the spring buds. In the winter bud of Abies firma, a temperatefir native to Japan, a low temperature exotherm was detectedaround –20?C, which is higher by 10?C than that of sub-alpineor sub-cold firs. The low temperature exotherms of these firsoccurred at nearly the same temperatures that result in thedeath of these primordial shoots. On the other hand, littleor no low temperature exotherm was detected in the winter budsof sub-cold spruces. In larch winter buds, numerous small exothermswere observed, which are probably due to the many leaf primordiain the buds. Unlike many temperate deciduous broad-leaved trees,no low temperature exotherm was detected below –15?C inwinter twig xylem of conifers such as Abies, Picea, Pinus, Larixand Pseudotsuga. Thus, very hardy coniferous twigs can tolerateextracellular freezing to –70?C. ¹ Contribution No. 1907 from the Institute of Low TemperatureScience. (Received June 8, 1978; )     

Hatchling painted turtles (Chrysemys picta) survive exposure to subzero temperatures during hibernation by avoiding freezing

Hatchling painted turtles (Chrysemys picta) were placed individually into artificial nests constructed in jars of damp soil and then were cooled slowly to temperatures between-7.7 and-12.7 °C. Distinct exotherms were recorded in all jars when water in the soil began to freeze at temperatures between-0.9 and-2.4 °C. A second (animal) exotherm was subsequently detected in some of the jars when water in hatchlings also began to freeze. An animal exotherm occurred in the temperature records for all 23 hatchlings that died in tests terminating at temperatures between-7.7 and-10.8 °C, but no such exotherm was apparent in the temperature records for the 23 turtles that survived these treatments. Moreover, the 4 hatchlings that produced exotherms in tests terminating between-11.5 and-12.7 °C failed to survive, but 5 of 7 hatchlings that produced no exotherm in these tests also died. Thus, turtles that die at subzero temperatures above-11 °C apparently succumb to freezing when ice propagates across their integument from the frozen soil, but animals that die at temperatures below-11 °C generally perish from some other cause. These findings indicate that hatchling painted turtles overwintering inside their shallow, subterranean nests survive exposure to subzero temperatures by avoiding freezing instead of by tolerating freezing.     

女贞和冬青卫矛叶片低温下胞外结冰模式的热力学新证据

低温条件下植物组织的结冰模式, 即胞外或胞内结冰, 直接决定着细胞的生死。目前胞外结冰的直接证据很少, 尤其缺乏热力学证据。用高分辨率差热扫描结合显微观察分析了女贞(Ligustrum lucidum)和冬青卫矛(Euonymus japonicus)叶片在降温过程中的结冰放热现象, 发现2种植物胞外结冰的热力学和组织结构变化的新证据。2种植物的活叶片在冷却过程中均呈现双放热峰, 即双相结冰的特征; 而预先冰冻杀死的叶片和叶片组织提取液浸润的滤纸片在同样冷却过程中仅有1个快速的大单放热峰, 即单相结冰的特征。显微观察也显示, 结冰过程中活组织细胞间隙中形成大量的白色冰晶, 且细胞虽然脱水收缩但细胞内的有色溶液没有流失, 表现出胞外结冰的特征。实验结果为深入揭示植物的冰冻伤害机制提供了新证据和研究方法。     

Supercooling of xylem ray parenchyma cells in tropical and subtropical hardwood species

 The freezing behavior of xylem ray parenchyma cells in several woody species, Ficus elastica, F. microcarpa, Mangifera indica, Hibiscus Rosa-sinensis, and Schefflera arboricola, that are native to non-frost tropical and subtropical zones, was investigated by differential thermal analysis (DTA), cryo-scanning electron microscopy (cryo-SEM) and freeze-replica electron microscopy. Although profiles after DTA did not exhibit clear evidence of supercooling in the xylem ray parenchyma cells, electron microscopy revealed that the majority of xylem ray parenchyma cells in all of the woody species examined were supercooled to around –10°C upon freezing temperatures and were not frozen extracellularly. It seems likely that DTA failed to reveal the low temperature exotherm (LTE), that is produced by breakdown of supercooling in the xylem ray parenchyma cells as a consequence of the overlap between the high temperature exotherm and the LTE in each case. The xylem ray parenchyma cells in these woody species were very sensitive to dehydration, and supercooling had, to some extent, a protective effect against freezing injury. It is suggested that the capacity for supercooling of xylem ray parenchyma cells of tropical and subtropical woody species might be the result of inherent structural characteristics, such as rigid cell walls and compact xylem tissues, rather than the result of positive adaptation to freezing temperatures. The present and previous results together indicate that the responses of xylem ray parenchyma cells in a wide variety of hardwood species to freezing temperatures can be explained as a continuum, the specifics of which depend upon the temperatures of the growing conditions. Received: 24 January 1997 / Accepted: 13 May 1997     

Freeze Desiccation: a Second Mechanism for the Survival of Hydrated Lettuce (Lactuca sativa L.) Seed at Sub-zero Temperatures

Differential Thermal Analysis of hydrated lettuce cv. GreatLakes achenes using a rapid cooling rate (20 °C h^–1)produced two exotherms per achene. Both exotherms representedthe freezing of supercooled water. The high temperature exothermoccurred at –93 °C and was produced by freezing ofwater inside the pericarp but exterior to the endosperm. Thetemperature at which it occurred could be altered by the additionof nucleating agents. The low temperature exotherm produced by freezing of the embryooccurred at –162 °C and marked the death of the seed.Its temperature was not changed by the addition of nucleatingagents but its occurrence required the structural integrityof the endosperm. At low cooling rates (1 and 2 °C h^–1)low temperature exotherms were not recorded and samples removedat –25 °C had high viability. Slow cooling causeda redistribution of water within the seed whereby ice formingoutside the endosperm caused desiccation of the embryo and preventedits freezing. A mechanism is proposed, in terms of established supercoolingand nucleation theory, to explain the observed results and thevalue of freeze tolerance to the species in its natural habitatis discussed. Cooling rate, differential thermal analysis, freezing avoidance, Lactuca sativa L., lettuce, seed, supercooling, water migration