Fingerprinting 146 Chinese chestnut(Castanea mollissima Blume)accessions and selecting a core collection using SSR markers

Chinese chestnut is an important nut tree around the world.Although the types of Chinese chestnut resources are abundant,resource utilization and protection of chestnut accessions are still very limited.Here,we fingerprinted and determined the genetic relationships and core collections of Chinese chestnuts using 18 fluorescently labeled SSR markers generated from 146 chestnut accessions.Our analyses showed that these markers from the tested accessions are highly polymorphic,with an average allele number(N_(a))and polymorphic information content(PIC)of 8.100 and 0.622 per locus,respectively.Using these strongly distinguishing markers,we successfully constructed unique fingerprints for 146 chestnut accessions and selected seven of the SSR markers as core markers to rapidly distinguish different accessions.Our exploration of the genetic relationships among the five cultivar groups indicated that Chinese chestnut accessions are divided into three regional type groups:group I(North China(NC)and Northwest China(NWC)cultivar groups),group II(middle and lower reaches of the Yangtze River(MLY)cultivar group)and group III(Southeast China(SEC)and Southwest China(SWC)cultivar groups).Finally,we selected 45 core collection members which represent the most genetic diversity of Chinese chestnut accessions.This study provides valuable information for identifying chestnut accessions and understanding the phylogenetic relationships among cultivar groups,which can serve as the basis for efficient breeding in the future.

Effects of the <i>Castanea mollissima Blume</i>Shell Cultivation Substrate on the Yield and Nutritional Composition of <i>Pleurotus geesteranus</i>

Cottonseed hull substrates blended with different ratios of Castanea mollissima Blume shell were prepared and used for the cultivation of Pleurotus geesteranus. The effects of the chestnut shell content on the mycelial growth rate, yield, nutritional composition and contents of heavy metals of the cultivated Pleurotus geesteranus were investigated. The results suggest that the Castanea mollissima Blume shell in substrate can increase the mycelial growth rate, yield, biological efficiency and the contents of protein, crude fiber, amino acids and essential amino acids of Pleurotus geesteranus. Further investigation suggests that the tannin and saponin in Castanea mollissima Blume shell and the C/N ratio of substrate significantly influence the mycelial growth rate. The crude fat content of Pleurotus geesteranus decreased, while the contents of heavy metals including mercury (Hg), arsenic (As), cadmium (Cd) and lead (Pb) increased with the increase of the Castanea mollissima Blume shell content in substrate. Based on these results, the content of Castanea mollissima Blume shell was optimized to be less than 30% for the cultivation of Pleurotus geesteranus.

板栗(Castanea mollissimaB1.)空苞形成因子的研究——Ⅱ.板栗空苞的胚胎学

选连续三年发生空苞率达90%的树为材科,对其胚眙发育进程进行了观察,并与连续三年不发生空苞树作比较。结果表明:板栗空苞发生的原因在胚胎学上有如下几方面,大孢子母细胞发育停止,减数分裂不能正常进行;成熟胚囊结构异常,卵细胞、助细胞解体;传粉受精不良导致受精作用异常;原胚早期敗育、胚乳败育、胚发育中止及部分空苞中种子的子叶不发育等?鄄熘谢狗⑾?总苞及子房外形的大小与胚珠发育进程之间有密切关系;另在珠形胚时期,一个子房内发育胚珠与退化胚珠在外形大小上也有明显差別。

板栗(Castanea mollissima Bl.)的胚胎学研究 Ⅱ雌配子体的超微结构

受精前的助细胞是代谢活跃的细胞,具丰富的线粒体、高尔基体和质体,内质网大多为光滑型且库槽膨大,极少淀粉粒;助细胞向合点端的壁是不连续的。卵细胞的壁与助细胞的相似,合点端也是不连续的;卵细胞质中线粒体,高尔基体和内质网均较少,但游离核糖体很丰富。中央细胞除具大量各种细胞器外,质体中还常有小淀粉粒。反足细胞壁在与中央细胞间及各反足细胞间是间断的,并具大量胞间连丝,细胞质中也富含各种细胞器,表明其代谢活动十分活跃;反足细胞中观察到核物质穿壁现象。珠心柱(postment)中部细胞在形态和结构上都与其外部细胞不同,显示了其功能的差异。文中并讨论了助细胞、反足细胞在胚囊营养中的作用以及胚囊营养物质供应的途径。

CTAB-silica Method for DNA Extraction and Purification from Castanea mollissima and Ginkgo biloba

A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method, the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour.

A new alkaloid from Castanea mollissima Blume

A new alkaloid, named 6-（2＇,3＇-dihydroxy-4＇-hydroxymethyl-tetrahydro-furan-1＇-yl）cyclopentadiene[c]pyrrole-1,3-diol, was isolated from the seeds of Castanea mollissima Blume. The structure was elucidated based on spectroscopic evidence including 2D NMR techniques.

板栗花发育B类基因CmPI的克隆及功能分析

【目的】PISTILLATA(PI)基因作为控制花器官发育的B类功能基因中的一员,在花器官发育中起到重要的作用。为探究PI基因在板栗花发育中的功能,对板栗花器官B类基因PISTILLATA(PI)进行鉴定和功能分析。【方法】在板栗基因组数据中检索PI同源基因CmPI,并克隆编码区序列。利用在线工具对CmPI进行生物信息学分析。通过荧光定量PCR分析CmPI在板栗不同组织及不同花发育时期的时空表达模式。构建35S::CmPI-GFP融合载体,瞬时转化烟草叶片,进行基因表达的亚细胞定位分析。将过表达载体35S::CmPI转入野生型拟南芥获得过表达植株,验证CmPI在花发育中的功能。【结果】克隆到的板栗CmPI基因开放阅读框长度630 bp,编码209个氨基酸,有高度保守的MADS域(MADS-box)和K域(K-box),属MADS的Ⅱ型亚家族转录因子,定位于细胞核。CmPI主要在板栗雄花中表达。在拟南芥中过表达CmPI导致萼片转变为花瓣样组织。荧光定量PCR结果表明,CmPI过表达拟南芥中除C类基因AtAG表达量显著高于野生型外,其他A类、B类、E类基因的表达量均低于野生型拟南芥。【结论】鉴定到的MADSbox基因CmPI为板栗花发育B类基因,可导致萼片花瓣化,可能是抑制雌花苞片进一步发育的关键基因。

罗田县板栗品种授粉配置研究

【目的】筛选湖北省罗田县板栗Castanea mollissima品种高产优质的授粉组合。【方法】选用‘桂花香’‘Guihuaxiang’、‘八月红’‘Bayuehong’、‘六月暴’‘Liuyuebao’、‘乌壳栗’‘Wukeli’、‘玫瑰红’‘Meiguihong’、‘浅刺大板栗’‘Qiancidabanli’等6个罗田县栽培的板栗品种,在花期相遇的基础上,设计22个授粉组合,开展品种授粉配置试验,对各授粉组合的授粉结实性和果实性状进行测定分析。【结果】异交显著降低了空苞率,除‘乌壳栗’外,各品种的异交空苞率均低于自交。异交显著提高了成果率(P<0.05),除‘乌壳栗’外,各品种的自交成果率均低于50%。授粉品种对各品种的刺苞性状、坚果性状、种仁的蛋白质和淀粉质量分数均有显著性影响(P<0.05)。综合成果率、果实性状等指标及主成分分析,筛选出各品种的成果率高、果实品质较好的授粉组合(♀×♂)是:‘桂花香’ב八月红’、‘八月红’ב六月暴’、‘六月暴’ב桂花香’、‘六月暴’ב八月红’、‘乌壳栗’ב六月暴’、‘玫瑰红’ב浅刺大板栗’、‘浅刺大板栗’ב桂花香’、‘浅刺大板栗’ב乌壳栗’。【结论】合理进行品种授粉配置可以有效降低罗田板栗品种的果实空苞率,提高果实产量与品质,实现高产优质栽培。

板栗(Castanea millissimaB1.)空苞形成因子的研究——Ⅰ.授粉受精对板栗空苞形成的影响

板栗空苞率高是其低产的重要原因之一。本研究通过人工授粉、剪除雌花柱头等试验发现:板栗不经授扮,座苞率高达68%以上;授粉受精不良不是板栗落果的主要原因,但不经授粉所结果球全为空苞。如选择亲和力强的花粉授扮,可以大大减少空苞;自花授粉者空苞率较高(平均高达18.3%),以混合花粉授粉,不仅空苞率很低(平均为0.4%),而且总苞内的果实数显著高于自花授扮。上述结果说明,授粉受精不良是板栗形成空苞的主要原因之一。

板栗黄化皱缩病检测方法优化及10个板栗品种抗性鉴定

为优化板栗黄化皱缩病检测方法,明确10个板栗品种的抗性,通过对巢式PCR引物添加量、退火温度的优化,结合植原体检出率和田间病情指数进行抗性评价。结果表明,巢式PCR在引物添加量为1.0μL、退火温度为54℃时,板栗植原体检出率最稳定;在毒源树嫁接400 d后,供试板栗品种均检测出植原体,其中‘燕山早丰芽变’叶片中植原体检出率最高,发病情况最严重,‘燕丽’叶片中植原体检出率最低,发病情况最轻;相关分析表明,植原体检出率和田间病情指数呈极显著正相关,相关系数为0.907,其线性回归方程为Y=0.276X+1.610;根据2种评价方法将供试板栗品种分为高抗、中抗和高感品种,其中‘燕秋’‘燕紫’‘燕龙’‘燕丽’为高抗品种,‘燕山早丰芽变’为高感品种。研究初步明确了10个板栗品种对黄化皱缩病的抗性差异,为选用抗性品种提供了基础数据。

栗实象成虫触角感器超微结构观察

栗实象Curculio davidi是为害板栗Castanea mollissima的主要种实害虫,严重影响着板栗的品质和产量。为探明栗实象成虫触角感器类型、形态和分布,本文利用扫描电镜对栗实象雌雄成虫触角及触角感器进行观察。结果表明:栗实象雌雄成虫触角柄节、梗节、鞭节1~6亚节长度存在显著差异。雌、雄成虫触角上均有7类12种感器,包括4种刺形感器、2种锥形感器、1种叉形感器、2种毛形感器,1种B9hm氏鬃毛、1种腔形感器和1种端指形感器。其中雄虫刺形感器Ⅰ、毛形感器Ⅰ、叉形感器的长度显著长于雌虫,雄虫鞭节第7、8亚节的锥形感器数量显著多于雌虫。

红色板栗和绿色板栗叶片光合特性及色素含量比较

以红色板栗红栗2号和绿色板栗燕宝为材料,通过Li-6400便携式光合仪测定叶片光合参数日变化以及红色板栗与绿色板栗的光响应曲线,同时使用紫外可见分光光度计测定叶片色素含量,探究红色板栗与绿色板栗叶片光合特性差异。结果表明:两种颜色板栗净光合速率日变化有明显区别,绿色板栗净光合速率为双峰形曲线,13:00时出现“光合午休”现象,而红色板栗为单峰形,无明显“光合午休”现象;红色板栗气孔导度、蒸腾速率、水分利用率日均值均大于绿色板栗,而胞间二氧化碳浓度日均值小于绿色板栗。光响应曲线结果表明,红色板栗有更高的最大净光合速率、光补偿点、暗呼吸速率、光饱和点与表观量子效率,与绿色板栗差异显著。对叶片色素含量进行分析,红色板栗与绿色板栗类胡萝卜素含量无明显差异,但是叶绿素含量和花色素苷含量差异显著。相关性分析结果表明,光响应特征参数中最大净光合速率、光饱和点与花色素苷含量有极显著正相关性,而光补偿点、暗呼吸速率与叶绿素含量呈现极显著或显著负相关。

气候变化下板栗适宜性分析及分布预测

【目的】探究影响板栗分布的主导环境因子,模拟不同气候条件下板栗潜在分布区,为板栗种质资源保护和引种提供理论依据。【方法】基于472个板栗现有种群分布地理信息与21个环境变量,采用物种分布组合模型模拟板栗潜在适宜分布区,通过AUC与TSS值评价模型准确性。利用组合模型推测末次冰盛期、全新世中期、当前、2030年和2050年板栗适宜区分布及其变化趋势。【结果】1)10种模型中,RF、GBM、MARES和GAM预测精度最好且稳定,最终组合模型AUC和TSS值分别为0.974和0.816,精度较高。2)影响板栗地理分布的主要环境因子为最冷月极端低温、海拔、最湿季降水量、年温变化范围和最湿月降水量,累计贡献率达89.79%。3)当前气候条件下中国潜在板栗适宜区分布广泛,自末次冰盛期以来,板栗适宜区不断向高纬度地区扩张,气候越暖扩张越快。但板栗整体适宜性不断下降,南方较为突出,北方则降幅较小,高适宜区总面积逐步减少。【结论】气候和地形因子共同影响板栗的地理分布,其中气候因子中温度因子贡献最大,地形因子中海拔因子贡献最大。当前气候条件下板栗在中国适宜区分布广泛,建议在高适宜区进行引种栽植及种质资源保护,当未来气候不利于板栗生长时向高纬度适宜区进行迁移引种,减少气候变化带来的损失。

气力式板栗采收装备悬浮速度的测定及影响规律

【目的】研究形状系数、含水率以及粒径对板栗悬浮速度的影响规律并建立不同粒径下板栗悬浮速度预测模型。【方法】以河北省唐山市迁西县的燕山早丰板栗为研究对象,通过自制悬浮速度试验台测量板栗的悬浮速度,并选取含水率、粒径、形状系数为影响因子,以悬浮速度为目标值,运用Origin软件进行单因素试验分析,建立不同粒径下板栗的悬浮速度预测模型,探究各参数与悬浮速度之间的变化规律。【结果】单因素试验中,粒径、含水率与悬浮速度呈现正相关,而形状系数与悬浮速度呈现负相关。不同粒径下板栗悬浮速度的变化范围为9.39 m/s~24.37 m/s;不同含水率下板栗悬浮速度的变化范围为12.78 m/s~23.12 m/s;不同形状系数下板栗悬浮速度的变化范围为9.18 m/s~24.84 m/s。【结论】本研究设计的板栗悬浮速度预测模型可以计算出板栗的最佳收获时期,降低设计气力输送装置时风机的功耗,提高板栗采收的效率,并为板栗气力输送、清选以及分级等机械设备的设计研究提供参考。

封开县油栗产业发展概况与对策建议

基于油栗(Castanea mollissima Blume.)相关文献资料和历年统计数据,结合实地调研的方法,对封开县油栗产业进行了综述。分析了封开县及其油栗产业的发展概况,厘清了封开县油栗产业发展中存在的问题,提出了精细化油栗种植管理、优化油栗种植结构及种植面积、强化果园配套设施建设、培育壮大龙头企业、加强一二三产业融合的对策与建议。

Ophiognomonia castaneae协同板栗褐缘叶枯病病原菌致病作用的研究

为明确板栗新病害褐缘叶枯病病原菌Phomopsis castaneae-mollissimae(Y1)与其共生菌板栗蛇孢日规壳Ophiognomonia castaneae(Y6)的关系,采用在发病不同时期组织分离、定期采样观察、致病性测定及对峙培养等方法进行研究。结果表明,发病初期在褐缘叶枯病病健交界处只分离到Y1,随病斑扩展Y6的分离频率逐渐增加,发病后期Y6的分离频率略高于Y1;Y6的无性型和有性型子实体均生长在病斑上;离体叶片接种Y1易产生病斑,接种Y6很少产生病斑,两菌混合接种比单独接种产生的病斑症状明显且发病速度快,表现出Y1致病力强,Y6致病力弱,Y6具有协同Y1致病的作用;Y6和Y1的营养体表现出明显的完全的亲和力。

基于LF-NMR探究板栗失水过程中质构和色差变化

板栗(Castanea mollissima Blu.)果实中的水分组成在其生理代谢及品质变化过程中具有重要作用。本研究以贵州黔西南望谟县板栗为实验材料,利用低场核磁共振技术,对不同水分含量(48%、38%、28%、18%)的板栗果中氢核的横向弛豫衰减信号进行测试,并利用SIRT模型对数据进行反演,得出横向弛豫时间T2的反演数据。采用自旋回波脉冲序列对板栗果的质子密度加权成像;使用物性测试仪分析质构特性,利用色差仪分析板栗失水过程中果肉颜色的变化。结果显示,自然失水改变了板栗果中水分的结合状态,水分发生明显迁移,自由水和不易流动水的信号幅值随失水程度的加剧而逐渐减弱。不同含水率板栗的反演峰总面积不同,随着含水率的增加,核磁共振信号增强,峰面积增大,两者具有显著的相关性。核磁共振成像技术通过图片明暗不同,可直观反映不同阶段的水分含量。板栗果实自然贮藏过程中发生石灰化,果实质地发生硬化,挤压咀嚼时抵抗力增大,失水初期内聚性和弹性变化不显著,后期下降;粘附性前期降低,但后期变化不明显;板栗失水初期果肉发生氧化,颜色逐渐呈暗红色,失水后期石灰化较为严重,果肉颜色变白,亮度增加。本研究结果可为板栗加工过程中的水分迁移机制探讨以及产品的品质控制提供参考。

Study on Effect of Different Varieties Configuration upon Yield of Yan Mountain Chestnut

In order to improve the yield and quality of 5 new Castanea mollissima varieties “Zipo”, “Zunyu” and traditional Yan Mountain Castanea mollissima cultivars “Duanci”, “Yanhong” and “Donglingmingzhu”, we carried out an experiment to study effect from different varieties configuration on yield of Yan Mountain chestnut. The result indicated that: cross-pollination had a significant impact on both fruiting rate and seed-setting rate;cross-pollination improved the nut number in each burr of each variety, in which “Duanci” was the biggest beneficiary, whose number of nut per burr increased from 1.08 to 2.77, 156% higher than that of the self-pollination combination;nut weight showed no significant difference in Xenia Effection;“Zunyu” improved the yield of “Yanhong” most;“Donglingmingzhu” was regarded as the best tree to improve “Zunyu”’s yield;the best variety to improve yield of “Zipo” was “Zunyu”;“Donglingmingzhu” and “Duanci” improved yield of each other most.

FTIR and Thermogravimetric Analysis of Three Kinds of Nutshells

The main components and pyrolysis characteristics of Camellia oleifera Abel hells, Castanea mollissima Blume shells, and Castanea mollissima Blume shells were analyzed by using FTIR and thermogravimetric methods. The experimental results indicated that the main components of the three kinds of raw materials consisted of cellulose, hemicellulose, and lignin. The highest contents of hemicellulose, cellulose, and lignin were in Camellia oleiferaAbel shells (49.34% ± 0.07%), Castanea mollissima Blume shells (27.34% ± 0.01%), and Carya cathayensis Sarg shells (49.78% ± 0.01%), respectively. The pyrolysis processes of three kinds of shells generally included three stages, namely dehydration, pyrolysis, and carbonization. The peak values and the appearance times of their pyrolysis rates were closely related to their compositions.

基于高光谱成像技术的板栗产地溯源

目的建立了一种基于高光谱成像(hyperspectral imaging,HSI)技术的板栗产地溯源模型。方法采集怀柔、迁西和沂蒙短枝3种不同产地板栗的高光谱图像,提取感兴趣区域,建立支持向量机(support vector machine,SVM)板栗产地溯源模型,通过比较分析不同预处理方法对建模结果的影响,选出最佳的预处理组合方法,并使用遗传算法(geneticalgorithm,GA)对模型进一步优化。结果实验结果表明,经多元散射校正(multiplicative scatter correction,MSC)和移动窗口平滑法(moving window smoothing,MWS)组合预处理后的数据所建立的溯源模型预测性能最好,分类的预测精确率达到了95%以上,模型整体的预测准确率为96.61%。经GA对SVM的参数C进行优化,优化后的模型对怀柔板栗和沂蒙短枝板栗的预测精确率达到了100%,模型整体的准确率提高到了98.31%。结论本研究基于高光谱成像技术建立了一种板栗产地溯源模型,经预处理和参数优化后,所建立的模型具有较好的预测性能,为板栗的产地溯源提供了一种新方法。