遮荫条件下乌拉苔草(Carex meyeriana)蒸腾特性及其与环境因子的关系

通过对不同遮荫下条件乌拉苔草蒸腾特性的研究表明 ,乌拉苔草蒸腾速率日变化规律是 ,上午 8h出现次峰 ,18h出现高峰 ,日变化有“午休”现象 ,其中光1和光3 出现在中午 12h ,对照和光2 出现在下午 14h。在上午8h ,蒸腾速率的变化不受气孔开闭的影响 ,是由于时间和环境因子的综合影响所致 ;中午出现的“午休”现象 ,是由于气孔关闭所至。不同遮荫处理下蒸腾速率变化规律基本一致 ,遮荫 5 0 %对蒸腾速率影响较大 ,其他处理对蒸腾速率的影响较小 ,说明乌拉苔草的蒸腾特性不仅由本身的生理特性所决定 ,并受环境因子综合的影响 ;另外 ,不同遮荫对气孔也有直接的影响 ,遮荫是影响气孔阻力的直接因素。

长白山区沟谷沼泽湿地乌拉苔草(Carex meyeriana)地上生物量与土壤有机质和氮素相关性分析

通过对长白山区沟谷沼泽湿地乌拉苔草群落的乌拉苔草生物量与该土壤有机质和氮素进行统计分析,揭示了该湿地上生物量与土壤有机质及氮素在土壤空间的异质性。相关分析结果表明,地上生物量和土壤有机质的相关系数各层均为不显著,但相关系数逐渐由小到最大,存在明显的空间差异;地上生物量与氮素之间的相关差异明显,但相关性均不显著;与硝态氮呈极显著和显著相关,但差异不大;与土壤中铵态氮的相关性均不显著,随着土壤剖面的加深相关系数增加。与全氮和速氮的相关性空间差异性很大。地上生物量和有机质、全氮、铵态氮、硝态氮相关性最大的是土壤剖面的C层,和速氮相关性最大的是表层。乌拉苔草生物量与土壤有机质和氮素在不同土壤剖面的相关性不同,揭示出该湿地有机质和氮素的空间异质性。

长白山区沟谷乌拉苔草Carex meyeriana沼泽湿地气候效应

通过对长白山区沟谷沼泽典型乌拉苔草湿地土壤温度、乌拉苔草群落相对湿度、叶片温度、风速、光量子通量密度以及蒸腾速率的日变化和蒸腾速率的季节变化的研究，初步揭示沟谷乌拉苔草沼泽湿地的气候效应。结果表明长白山沟谷沼泽湿地具有三江平原沼泽同样的冷湿效应。土壤化通时间比三江平原化通时间提前1个多月，蒸腾速率日变化不同季节趋势基本一致，不同层次叶片的蒸腾速率日变化趋势基本一致，最大值在7至8月份蒸腾速率（H2O）达到100～140mol·m^-2·S^-1。光量子通量密度与温度成正比，与湿度成反比，大气温度日变化和叶片温度日变化趋势基本相同，相对湿度的日变化趋势和大气温度的变化趋势正好相反，和光量子通量密度的变化趋势相反。沟谷湿地的同样具有小气候效应。

长白山区沟谷沼泽乌拉苔草(Carex meyeriana)湿地土壤酶活性与氮素、土壤微生物相关性研究

通过现场采样及室内分析,研究了长白山区沟谷沼泽乌拉苔草湿地土壤酶活性及其与氮素、土壤微生物的相关性。结果表明,土壤脲酶活性在时空变化中,表层最高,且最大值出现在6月份,为2.57;C层最小,最小值出现在7月份,为0.3;土壤纤维素酶活性表层最高出现在6月份,为1.35;B层最小,最低点出现在6月份,为0.18,土壤蛋白酶活性在时空变化中变化规律基本一致,最高值是6月份B层,为8.5;最低点是C层的5月,为0.9;与氮素的相关性分析结果为:土壤脲酶在8月份最大,为0.91,B层最大为0.76;土壤纤维素酶在5月份最大,为0.41;C层最大,为0.52;土壤蛋白酶在4月份最大,为0.83;A层最大,为0.67,均不呈显著相关。与微生物相关结果为:土壤脲酶与8月份微生物呈极显著的正相关;蛋白酶与B层的细菌呈显著的正相关;脲酶与7、8月份的放线菌呈极显著的正相关。本文揭示了土壤不同酶活性受制于不同的微生物影响。

云南疣粒野生稻Oryza meyeriana Baill.总黄酮含量分析研究

疣粒野生稻(Oryza meyerianaBaill.)在云南部分地区被用作消炎草药,这种利用行为对该野生稻的生存造成了一定的影响和威胁,为了有效地保护野生稻,对疣粒野生稻是否含有消炎药用成分——总黄酮含量进行了分析。通过本研究,建立了测定疣粒野生稻总黄酮含量的分光光度法,测定了云南省疣粒野生稻2个群体31个居群及居群内单株的总黄酮含量。结果表明疣粒野生稻总黄酮平均含量低于对照银杏叶和竹叶;原生境群体总黄酮含量(1.158g/kg)高于温室群体含量(0.928g/kg),但t测验差异不显著。探讨了云南疣粒野生稻作为消炎草药的可能性和可行性,提出在重要种质资源保护和利用过程中应重视土著知识的利用。

ISSR Analysis on Genetic Diversity of the 34 Populations of Oryza meyeriana Distributing in Yunnan Province,China

The genetic diversity of the 34 populations of wild rice Oryza meyeriana Baill. distributed in Yunnan Province, China was analyzed using 13 inter-simple sequence repeat （ISSR） markers. A total of 168 bands were amplified, of which 135 polymorphic bands were discovered and the percentage of polymorphic bands （PPB） was 80.36%. A genetic diversity was revealed as Nei＇s gene diversity （H） = 0.2666 and Shannon information index （I） = 0.4028 at population level. The 34 populations were divided into different groups based on administrative regions, latitude and longitudes, river areas, altitudes of their origins, and their indexes such as Na （number of alleles）, Ne （effective number of alleles）, H, I and PPB were calculated. Richer genetic diversity was found in the wild rice populations distributed in Simao Prefecture than that in Lingcang Prefecture or Xishuangbanna Prefecture whereas the least genetic diversity was in Baoshan Prefecture or Dehong Prefecture. Rich genetic diversity was also discovered in the wild rice populations originated from higher than 710 m altitude around the middle and lower reaches of the Lancang River belonging to the Pacific Ocean drainage system. The 34 populations could be classified into two groups, one group covered the wild rice distributing in Simao Prefecture only while the other group covered ones in Lingcang, Xishuangbanna and Dehong Prefectures. The issue on how to effectively conserve the wild rice germplasm was discussed.

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L.

Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O.sativa and O.meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance.

Comparative Analysis of Genomes in Oryza sativa, O. officinalis, and O. meyeriana with C_(0)t-1 DNA and Genomic DNA of Cultivated Rice

Fluorescence in situ hybridization （FISH） and comparative genomic hybridization （CGH） were applied to somatic chromosomes preparations of Oryza sativa, O. officinalis, and O. meyeriana with labeled probes of Cot-1 DNA and genomic DNA＇from the cultivated rice. The coverage percentage （%） and size （Mb） of Cot-1 DNA in O. sativa, O. officinalis, and O. meyeriana were 47.1 ±0.16, 38.61 ±0.13, 44.38＋_0.13, and 212.33 ± 1.21,269.42 ± 0.89, 532.56± 1.68 Mb, respectively. The coverage percentage and size of genomic DNA from O. sativa in O. officinalis and O. meyeriana were 91.0, 93.6% and 634, 1 123 Mb, respectively, in which 365 and 591 Mb in O. officinalis and O. meyeriana were from O. sativa genomic DNA, but not from repetitive sequences of O. sativa, and the uncoverage genome size in O. officinalis and O. meyeriana were 64 and 78 Mb, respectively. In addition, karyotype analysis was conducted based on the signal bands of Cot-1 DNA in O. sativa, O. officinalis, and O. meyeriana. The results showed that highly and moderately repetitive sequences in Oryza genus were conserved as the functional genes during evolution. The repetitive sequences reduplication may be one of the important causes of the genome enlargement of O. officinalis and O. meyeriana, and O. officinalis genome enlarged more slowly when compared with O. meyeriana. Based on the above results, it is concluded that O. officinalis and O. meyeriana were formed by reduplication, rearrangement, and gene selective loss during the evolution process.

长白山区沟谷乌拉苔草(Carex.meyeriana)沼泽湿地水文物理效应

通过对长白山区沟谷乌拉苔草（C.meyeriana）沼泽湿地和三江平原乌拉苔草沼泽湿地土壤物理性质和乌拉苔草根系生物量的测定，对水文物理效应进行了分析，结果表明：长白山区沟谷乌拉苔草沼泽湿地的土壤孔隙度小于三汀平原乌拉苔草沼泽湿地，容重和体积质量则高于三江平原乌拉苔草沼泽湿地。土壤蓄水能力低于三江平原乌拉苔草沼泽湿地；乌拉苔草发达的根系具有强大的蓄水能力．尤以根鞘的蓄水能力最强，蓄水可以占鲜质量的91．1％-93．72％，比根系蓄水能力高出2．54％-33．94％。由此得知乌拉苔草沼泽湿地的具有巨大的蓄水能力。是天然的水调节器。通过分析可知其水文物理过程对沼泽湿地对其上游河流具有调节作用，同时还可以保存上游河流生物多样性，以及严重的影响着水面以下化学过程的发生。

Transpiration Rates of Carex Meyeriana in Relation to Micrometerological Factors in a Mountain Valley Wetland

In order to reveal transpiration rates of wetland plants and its relationships to micro-meterological factors in a mountain valley wetland, relative humidity, air temperature, leaf temperature, soil temperature, photo flux density and transpiration rates were measured once two hours in a Carex meyeriana wetland of the Changbai Mountain valley in dry (July) and wet (September) of 2003, respectively. Results showed that the tendency of "decreasing after increasing" was obvious in wet season. However, a relatively stable trend was observed for the transpiration in dry season.. Generally, the photon flux density of Carex meyeriana was higher in wet season than that in dry season. However, the variabilities of leaf temperature, air temperature and relative humidity were similar in both seasons. Higher transpiration rates of Carex meyeriana leaves were observed in July (varied from 40 to 150 mol·m-2·s-1) compared to those (varied from 7 to 14 mol·m-2·s-1) in September. Transpiration rates were significantly correlated with air temperature (P<0.01), leaf temperature (P<0.01), and wind speed (P<0.05), but correlationship between relative humidity and photo flux density was not significant (P<0.05).

A study on population genetic structure of Oryza meyeriana (Zoll. et Mor. ex Steud.) Baill. from Yunnan and its in situ conservation significance

In order to determine genetic diversity of Oryza meyeriana (Zoll. et Mor. ex Steud. ) Baill., 12 enzyme systems encoded by 17 loci were electrophoretically analyzed in 164 individuals of seven populations from Simao Prefecture, Yunnan Province, China. In comparison with those seed plants with the same life history and breeding systems, as well as the other species in the genus Oryza, the species shows rather low levels of genetic diversity (A = 1.1, P = 8.0%, Ho = 0.004 and He = 0. 015) within populations and high genetic differentiation among populations. FST was up to 0. 649, suggesting that 64. 9% of total genetic variability exists among populations. Considering high genetic differentiation among populations from a limited geographic region, most of the populations of the species are worth being protected, and therefore, great natural protection regions should theoretically be established in which a great deal of populations should be involved for developing in situ conservation management. Meanwhile, some priory localities for in situ conservation of O. meyeriana in Yunnan Province, were proposed.

Crossability Barriers in the Interspecific Hybridization between Oryza sativa and O.meyeriana

Oryza meyeriana Baill （GG genome） is a precious germplasm in the tertiary gene pool of cultivated rice （AA genome）, and possesses important traits such as resistance and tolerance to biotic and abiotic stress. However, interspeciflc crossability barrier, a critical bottleneck restricting genes transfer from O. meyeriana to cultivars has led to no hybrids through conventional reproduction. Therefore, the reasons underlying incrossability were investigated in the present report. The results showed that： （i） at 3-7 d after pollination （DAP）, many hybrid embryos degenerated at the earlier globular-shaped stage, and could not develop into the later pear-shaped stage. Meanwhile, free endosperm nuclei started to degenerate at 1 DAP, and cellular endosperm could not form at 3 DAP, leading to nutrition starvation for young embryo development; （ii） at 11-13 DAP, almost all hybrid ovaries aborted. Even though 72.22% of hybrid young embryos were produced in the interspecific hybridization between O. sativa and O. meyeriana, young embryos were not able to further develop into hybrid plantlets via culturing in vitro. The main reason for the incrossability was hybrid embryo inviability, presenting as embryo development stagnation and degeneration since 3 DAP. Some possible approaches to overcome the crossability barriers in the interspecific hybridization between O. sativa and O. meyeriana are discussed.

Somatic cell cryopreservation and protoplast regeneration of important disease-resistant wild rice Oryza meyeriana Baill

Oryza meyeriana Baill is one of the three wild rice species found in China. O.meyeriana possesses valuable characteristics but is reluctant in cell culture in vitro . In a series of experiments, callus with no regeneration ability was induced from young panicle of O.meyeriana. The callus was subcultured and propagated. Embryogenic cell clones were obtained after cryopreservation. Suspension cultures were established and protoplasts were isolated and regenerated into plants. Results of artificial inoculation of Xanthomonas campestris pv. Oryzae showed that the strong resistance did not change in the regenerated plants. The development of protoplast to plant system is an important progress towards utilization of O.meyeriana via cellular engineering. The experiments demonstrated that cryopreservation of plant calli was a new way to obtain embryogenic cell line.

Identification of molecular markers linked to rice bacterial blight resistance genes from Oryza meyeriana

Y73 is a progeny of asymmetric somatic hybridization between Oryza sativa cv.Dalixiang and the wild rice species Oryza meyeriana.Inoculation with a range of strains of Xanthomonas oryzae pv.oryzae showed that Y73 had inherited a high level of resistance to rice bacterial blight(BB)from its wild parent.An F2 population of 7125 individuals was constructed from the cross between Y73 and a BB-susceptible cultivar IR24.After testing 615 SSR and STS markers covering the 12 rice chromosomes,186 markers were selected that showed polymorphism between Y73 and IR24.Molecular markers linked to the BB resistance genes in Y73 were scanned using the F2 population and the polymorphic markers.The SSR marker RM128 on chromosome 1,the STS marker R03D159 on chromosome 3 and the STS marker R05D104 on chromosome 5 were found to be linked to the rice BB resistance genes in Y73.

Transfer of bacterial blight resistance from Oryza meyeriana to O. sativa L. by asymmetric somatic hybridization

Asymmetric somatic hybrid plants were pro-duced between cultivated rice (Oryza sativa L.) and wild spe-cies [O. meyeriana (Zoll. etMor. exSteud.)] with high resis-tance to rice bacterial blight. X-ray-irradiated protoplasts of the wild species were used as donor and chemically fused with iodoacetamide-inactivated protoplasts of rice cv. 02428 to produce hybrids. Seventy-two plants were regenerated from 623 calli based on metabolic complementation. The morphological characters of the plants closely resembled that of the rice. Simple sequence repeats were employed to iden-tify their hybridity. Cytological analysis of root-tips revealed that their chromosome number varied in the range of 27— 38. The somatic hybrids were inoculated with strains of Xanthamonas oryzae pv. oryzae at adult growth stage and demonstrated the resistance to bacterial blight introgression from the O. meyeriana.

乌拉草提取物对大鼠皮肤烫伤后金黄色葡萄球菌感染的抑菌作用

目的:探讨乌拉草提取物黄酮类成分对大鼠皮肤烫伤后金黄色葡萄球菌(S.aureus)感染的抑菌作用,为新型抗感染中药的研发提供新思路。方法:采用水醇提取法提取乌拉草有效成分并利用紫外分光光度法测定其黄酮含量,采用牛津杯法抑菌试验检测乌拉草有效成分对细菌的抑菌活性,试管稀释法测定其最低抑菌浓度(MIC)。24只SD大鼠采用自制烫伤仪构建Ⅱ度烫伤模型,随机分为生理盐水对照组、青霉素钠(NaBP)对照组和乌拉草提取物CM-70治疗组,每组8只。3组大鼠创面皮内多点注射接种20μL(1.0×10^(8)CFU·mL^(-1))S.aureus,次日3组大鼠分别涂抹无菌生理盐水(0.5 g无菌脱脂棉吸附生理盐水0.5 mL·cm^(-2))、NaBP(0.5 g无菌脱脂棉吸附NaBP,5000 U·kg^(-1)体质量)和10 g·L^(-1)乌拉草CM-70提取物稀释液(0.5 g无菌脱脂棉吸附0.5 mL·cm^(-2)乌拉草CM-70提取液)。观察各组大鼠创面感染情况,采用创面痂下细菌定量实验检测各组大鼠烫伤后第1、2、3、4、5、6和7天创面细菌数(CFU·g^(-1)),酶联免疫吸附试验(ELISA)法检测各组大鼠烫伤前和烫伤后第1、3、7及14天血清白细胞介素1β(IL-1β)、白细胞介素10(IL-10)和肿瘤坏死因子α(TNF-α)水平。结果:体外抑菌活性检测,不同稀释浓度乌拉草CM-70提取物稀释液针对金黄色葡萄球菌形成不同大小抑菌圈,针对大肠埃希菌和白色念珠菌未形成抑菌圈。MIC检测,乌拉草有效成分对S.aureus的MIC值为2.5 g·L^(-1)。各组大鼠烫伤创面感染和愈合情况观察,与生理盐水对照组比较,感染后第3天,乌拉草CM-70提取物治疗组大鼠伤口浆液渗出较少;感染后第7天,伤口浆液渗出减少;红肿减轻,伤口面积缩小;感染后第14天,伤口创面已完全干燥,无浆液渗出,表面形成痂皮,伤口愈合良好,与NaBP对照组相近。烫伤创面痂下细菌定量,与生理盐水对照组比较,同一时间点乌拉草CM-70提取物治疗组大鼠创面痂下细菌数明显减少(P<0.01)。ELISA法检测,与生理盐水对照组比较,乌拉草CM-70提取物治疗组大鼠血清中IL-10水平在烫伤后第1和3天明显升高(P<0.05),在第7和14天明显降低(P<0.05);乌拉草CM-70提取物治疗组大鼠血清中IL-1β和TNF-α水平在烫伤后第1、3、7和14天明显降低(P<0.05)。结论:乌拉草有效成分可明显抑制大鼠皮肤烫伤后S.aureus感染,通过调控血清IL-1β、IL-10和TNF-α水平发挥抗炎和抑菌作用。

Box-Behnken设计优化乌拉草中总黄酮和总多酚的提取工艺研究

目的优化乌拉草中总黄酮、总多酚的提取工艺。方法乙醇回流法,以总黄酮和总多酚的含量为考察指标,采用单因素实验考查乙醇浓度,料液比、提取时间,采用响应面设计方法优化乌拉草中总黄酮、总多酚的提取工艺。结果在单因素考察基础上,通过响应面设计优化乙醇回流法提取乌拉草中总黄酮和总多酚的最佳工艺为:乙醇浓度80%;料液比1∶30;提取时间90 min。结论本方法提取乌拉草中总黄酮、总多酚的工艺合理,重现性良好,可以为乌拉草资源深度开发提供理论依据。

不同积水处理下乌拉苔草蒸腾速率日变化及其与环境因子的关系

在相同的光照、相对湿度、温度和风速等自然环境下,对相同土壤条件下的乌拉苔草湿地进行不同水分的处理,以桶(高50cm、直径40cm)中水分为基准,设为满水(干1)、半水(干2)、干旱(干3)3个处理,以自然状态下的土壤为对照。对乌拉苔草叶片的蒸腾速率、气孔阻力、光合有效辐射、空气温度、叶片温度、相对湿度、风速及大气中CO2的体积分数等指标进行了测定。结果表明:不同积水条件下,蒸腾速率的日变化与正弦曲线相似;气孔阻力的日变化呈现单峰形变化,但峰值不同,干2峰值最小,不同处理的峰值出现的时间相同。从时间上看,气孔阻力受水分的影响,与对照相比提前达到最大值。干1、干2、干3处理的蒸腾速率和气孔阻力呈显著的负相关,风速和蒸腾速率呈显著的正相关;干2、干3处理的光量子通量密度和大气温度呈显著的正相关;干3处理的蒸腾速率和光量子通量密度呈显著的正相关。

长白山沟谷湿地乌拉苔草沼泽湿地土壤微生物动态及环境效应研究

实验主要研究了乌拉苔草湿地土壤微生物不同季节在不同土壤剖面上的动态变化,结果表明:不同季节、不同土壤剖面细菌数量最大,其次是真菌,最少的是放线菌;细菌数量在表层和A层土壤中随着季节(6～9月份)变化不断增加,B层和C层7月份较低,从8月份开始上升直到9月份;放线菌在不同的剖面变化较大,在C层仅9月份有,数量极少;真菌在不同剖面变化一致,随着季节变化(6～9月份)不断增加。研究表明:乌拉苔草湿地土壤微生物数量的变化反映了该湿地本身特有的环境效应。

中国“疣粒野稻”拉丁学名和中名的订正

中国“疣粒野稻”的小穗近等长于颗粒野稻,而短于疣粒野稻。内外稃(谷壳)表面电镜扫描形态是:中国“疣粒野稻”的山形瘤状突起分布较密,与颗粒野稻近似,但疣粒野稻则分布较疏。中国“疣粒野稻“的钩毛多为弯锥形:钩毛周围的硅质突起为乳头状,顶端圆而光滑。疣粒野稻和颗粒野稻的钩毛为雀嘴形;钩毛周围的硅质突起为火山顶状,顶端具星状冠。以上述形态为主要依据,中国“疣粒野稻”与颗粒野稻和疣粒野稻均有明显区别;在地理分布方面也与上二亚种不同。所以,现将中国“疣粒野稻”另立一新亚种,为瘤粒野稻Oruza meyeriana (Zoll. et Mor.)Baill. subsp. tuberculata W. C. Wu et Y. G. Lu, G. C.