亚麻外源基因遗传转化研究进展

亚麻作为重要的经济作物,其遗传转化研究对亚麻新品种的选育工作具有重要意义.分析亚麻遗传转化方法、提高转化效率及转化组织筛选等方面,重点从抗除草剂基因、抗病基因、抗重金属逆性基因、改善亚麻纤维质量基因,以及改善亚麻油质量基因方面综述亚麻遗传转化外源基因的研究和应用进展,并讨论亚麻遗传转化过程中存在的问题和解决策略.     

亚麻EST序列中SSR标记的筛选

利用亚麻NCBI数据库中的7 941条亚麻EST序列进行SSR的筛选,共发现222个SSR,占整个EST数据库的2.73%,其中三核苷酸重复单元的EST-SSR占总SSR的72.1%,二核苷酸和四核苷酸二者出现的频率基本相近,分别占总SSR的14.4%和13.5%.AGAA是四核苷酸中的优势重复类型,占四核苷酸重复类型的67.67%.设计的21对EST-SSR引物中有18对在10个亚麻材料中有扩增产物,占设计引物的85%,有14对产物条带比较清晰并具有多态性.基于SSR标记进行聚类分析,可将10个亚麻材料划分为3个组.本研究建立的亚麻SSR标记,为亚麻遗传多样性鉴定、分子作图等研究提供了一种有效的分子标记系统.     

宁夏十字花科1种新记录植物

报道了宁夏十字花科1种新记录植物,小果亚麻荠(Camelina microcarpa).     

基于农艺性状和ISSR标记分析亚麻种源的变异及遗传关系

根据农艺性状的测定结果及ISSR标记分析结果,采用聚类分析方法对引种至云南昆明的73份亚麻(Linum usitatissimum L.)种源的变异及遗传关系进行了研究.结果表明,73份亚麻种源的10个农艺性状(包括株高、工艺长度、茎粗、单株分枝数、单株蒴果数、单株茎干质量、单株种子产量、单株纤维产量、种子千粒重和出麻率)均有一定的差异,但总体上生长状况均较好;根据农艺性状可将73份亚麻种源分成2类,每一类又可分为2组,其中,第Ⅰ类包括36份种源,第Ⅱ类包括37份种源.从40个ISSR引物中筛选出9个引物用于ISSR扩增,共扩增出54条带,其中多态性条带33条,多态性条带百分率为61.1%;基于ISSR扩增结果进行聚类分析,在相似系数0.80处可将供试的亚麻种源分成3大类:第1类包括67份种源,第2类包括4份种源,第3类仅有2份种源,在相似系数为0.82处第1类种源又可分成3个亚类.结果显示,根据农艺性状和ISSR标记进行的聚类分析结果有一定差异,但对二者进行综合分析,结合亚麻不同种源的适应性及育种目的,可培育出适于本地栽培的优良杂交后代.     

亚麻胚性细胞悬浮培养体系的建立和影响因素

亚麻品种‘双亚五号’的胚性愈伤组织诱导最佳培养基为MB（MS无机盐加B5维生素）＋1.0mg·L^-12,4-D;细胞初始悬浮培养的最佳培养基为MB＋0.2mg·L-16-BA;细胞继代悬浮培养的最佳培养基为改良的MB（NH4NO3减半,KNO3加倍）＋0.02mg·L^-12,4-D＋0.2mg·L^-16-BA;适宜的蔗糖量为50g·L^-1;适宜的继代接种量为1.0～1.5g·（25mL）^-1;继代间隔为5～7d。     

亚麻SRAP反应体系的优化

通过研究亚麻SRAP反应体系中主要因子对扩增结果的影响,建立了亚麻SRAP-PCR反应的优化体系.在20μL的反应体系中将PCR的5个主要成分分别设定8个浓度梯度,结果表明,最适宜的优化浓度分别为:1.5 mmol/L Mg2+、0.3 mmol/L dNTP、1.5 U Tap酶、30 ng/μL模板DNA 90 ng和25 ng/μL引物100 ng.用6个亚麻材料验证优化体系,检测结果显示,多态性高,反应体系的稳定性和可重复性好,为SRAP标记技术在亚麻分子生物学研究方面的应用奠定了基础.     

Characterization of Interspecific Hybrids Between Oryza sativa L. and Three Wild Rice Species of China by Genomic In Situ Hybridization

In the genus Oryza, interspecific hybrids are useful bridges for transferring the desired genes from wild species to cultivated rice （Oryza sativa L.）. In the present study, hybrids between O. sativa （AA genome） and three Chinese wild rices, namely O. rufipogon （AA genome）, O. officinalis （CC genome）, and O. meyeriana （GG genome）, were produced. Agricultural traits of the F1 hybrids surveyed were intermediate between their parents and appreciably resembled wild rice parents. Except for the O. sativa × O. rufipogon hybrid, the other F1 hybrids were completely sterile. Genomic in situ hybridization （GISH） was used for hybrid verification. Wild rice genomic DNAs were used as probes and cultivated rice DNA was used as a block. With the exception of O. rufipogon chromosomes, this method distinguished the other two wild rice and cultivated rice chromosomes at the stage of mitotic metaphase with different blocking ratios. The results suggest that a more distant phylogenetic relationship exists between O. meyeriana and O. sativa and that O. rufipogon and O. sativa share a high degree of sequence homology. The average mitotic chromosome length of O. officinalis and O. meyeriana was 1.25- and 1.51-fold that of O. sativa, respectively. 4＇,6＇-Diamidino- 2-phenylindole staining showed that the chromosomes of O. officinalis and O. meyeriana harbored more heterochromatin, suggesting that the C and G genomes were amplified with repetitive sequences compared with the A genome. Although chromocenters formed by chromatin compaction were detected with wild rice-specific signals corresponding to the C and G genomes in discrete domains of the F1 hybrid interphase nuclei, the size and number of O. meyeriana chromocenters were bigger and greater than those of O. officinalis. The present results provide an important understanding of the genomic relationships and a tool for the transfer of useful genes from three native wild rice species in China to cultivars.     

亚麻酶法脱胶研究进展

该文综述了亚麻酶法脱胶的研究现状及前景。从酶的组成、酶法亚麻脱胶工艺和酶法脱胶后亚麻纤维结构及组成变化三方面进行了论述。指出了亚麻酶法脱胶的优越性,现存的问题及其研究方向。     

尖孢镰刀菌亚麻专化型Folprp4基因参与调控菌丝生长和分生孢子发生

目的: 通过对尖孢镰刀菌中Folprp4基因的鉴定,揭示其在尖孢镰刀菌中的功能及致病相关性。方法: 基于同源重组原理,根据测定出的Folprp4基因序列,应用Split-Marker重组技术构建含有潮霉素抗性基因(hph)的基因缺失盒。将基因缺失盒经PEG介导转化到野生型原生质体中,在含有潮霉素B的TCC培养基上筛选转化子,通过PCR正负筛查获得Folprp4基因缺失突变株(ΔFolprp4)。构建含有Folprp4基因的载体pZDH1,并将其转化到敲除突变体中进行互补测验。结果: 与野生型(hm)和异位插入突变体(ecFolprp4)相比,敲除突变体菌丝生长受到严重阻碍,当野生型和异位插入突变体长满整个平板时,敲除突变体菌落呈小点状。敲除突变体的另一个显著变化是ΔFolprp4的分生孢子产量显著下降。侵染实验表明,ΔFolprp4对亚麻幼苗的毒力显著降低。互补实验表明,该互补载体的回复子(Folprp4-C)在菌落形态、生长速率、分生孢子产量和毒力方面均恢复到了野生型菌株。结论: Folprp4基因与尖孢镰刀菌的菌丝生长、分生孢子发生和致病性有关。     

Eco-physiological responses of linseed (Linum usitatissimum) to salt and alkali stresses

AimsEffects of salt and alkali stresses (NaCl-Na₂SO₄ and NaHCO₃-Na₂CO₃) were compared on growth, photosynthesis characters, ionic balance and osmotic adjustment of linseed (Linum usitatissimum), to elucidate the mechanisms of salt and alkali stress (high pH value) damage to plants, and their physiological adaptive mechanisms to the stresses. MethodsThe experiment was carried out in an artificial greenhouse. Plants grew at approximately 700 mmol·m^-2·s^-1 photosynthetic photon flux density (PPFD) in greenhouse under photoperiod of 15 h in light and 9 h in dark. In each plastic pot (17 cm diameter) which contained 2.5 kg of washed sand, 20 linseed seeds were sown. The seedlings were exposed to stresses lasting 14 days after 2 months.Important findingsThe inhibitory effects of alkali stress on linseed growth were more remarkable than those of salt stress, indicating that alkali and salt represent two distinct forms of stress. The alkali stress increased the Na⁺ content in shoots, damaged the photosynthetic system, and highly reduced the net photosynthetic rate and C assimilation capacity. Under salinity stress, the Na⁺ content increased, the K⁺ content decreased with increasing stress. Greater changes were observed under alkali than under salt stress. Alkali stress caused the massive influx of Na⁺, which probably explained that the harmful of alkali stress on plants was stronger than that of salt stress. Under alkali stress, Ca²⁺ and Mg²⁺ decreased in roots, showing that high pH value around roots hindered the absorption of them. Fe²⁺ and Zn²⁺ had little effects on the osmotic adjustment, mainly because of they had a low ion content. Under salt stress, anion increased in order to balance the sharp increase of Na⁺. However, alkali stress made severe deficit of negative charge, broke the intracellular ionic balance and pH homeostasis, and caused a series of strain response. Our results showed that linseed enhanced the synthesis of soluble sugars to balance massive influx of Na⁺ under salt stress, but linseed enhanced the synthesis of organic acids to compensate for the shortage of inorganic anions, which might be a key pathway for the pH adjustment. In conclusion, the alkali stress (high pH value) clearly inhibited the growth, element absorption, ion homeostasis reconstruction of plants. Organic acid concentration is possibly a key adaptive factor for linseed to maintain intracellular ion balance and regulate high pH value under alkali stress.