萝卜花粉的营养成分研究

萝卜花粉的营养成分测定表明,其蛋白质含量28.38%,总糖含量33.33%,氨基酸总量27.59%,总黄酮含量3.57%,维生素和矿质元素含量亦较丰富。萝卜花粉资源丰富,营养成分齐全,具有很好的开发利用价值。     

萝卜溶菌酶诱导条件的研究

分别以不同浓度（Ａ_（４５０ｎｍ）表示）的高压灭活白色葡萄球菌、大肠杆菌及水杨酸对萝卜溶菌酶进行不同时间的诱导．结果表明,诱导时间２４ｈ效果最好,白色葡萄球菌最适浓度Ａ_（４５０ｎｍ）为０．９４,酶的活力提高了２．３４倍;大肠杆菌最适浓度是Ａ_（４５０）ｎｍ为０．５５,诱导酶活力提高了０．７５倍,以０．２％的水杨酸为诱导物时,第４ｄ和第８ｄ的效果最好,酶活力分别提高了２．４和１．７倍．以酶作用产物进行田间叶面喷施和叶面喷施加根部灌施进行诱导,４８ｈ后酶活力提高２．１６和３．６６倍．     

萝卜低温胁迫转录因子的克隆及遗传转化

采用RT-PCR和电子克隆技术从抗寒植物萝卜(Raphanus sativusL.)中分离了一个低温胁迫转录因子的cDNA全长序列,命名为RsICE1(GenBank登录号为HQ891287).序列分析显示,该基因全长1 375 bp,编码区为1266 bp,编码421个氨基酸.序列比对表明,该蛋白C端含有一个典型的bHLH结构域,与其他植物的ICE1蛋白具有较高的同源性.进化树分析表明,RsICE1编码的蛋白与油菜的ICE1编码蛋白亲缘关系最近,处在同一进化分枝.半定量RT-PCR分析表明,RsICE1是冷诱导条件下差异表达的基因.构建该基因的植物表达载体,利用农杆菌介导法转化粳稻品种龙粳24,经PCR和RT-PCR分子检测,证明RsICE1基因已经整合到水稻基因组中,并正常表达.与对照相比,低温处理后转基因株系存活率和脯氨酸含量明显增加,相对电导率积累速率明显下降,提高了抗低温胁迫能力.     

Non-Random Mating in a Wild Radish, Raphanus sativus

Abstract When mating is non-random among several, compatible donors, the fitness of pollen donors, maternal plants, and offspring may be affected. Although this process may be important, it is much less studied than other forms of non-random mating such as incompatibility and avoidance of inbreeding. Therefore, the amount and consequences of non-random mating were investigated in greenhouse studies with wild radish, Raphanus sativus . Six compatible donors differed in the number, position, and weight of seeds sired, so mating was non-random at the level of mate identity. Mate number also affected mating patterns; fruits with more fathers were allocated more resources. This keeps mate number per fruit high. In contrast, other processes appear to keep mate number below the maximum so that mate number per fruit is regulated at an intermediate level. Mate identity had clear consequences as offspring with different fathers were of different sizes after 11 weeks. The effects of mate number on offspring success were less clear. These and other data suggest that non-random mating among compatible donors is a relatively common process in wild radish. It may occur through mechanisms controlled by the pollen tubes, the maternal plants or the embryos. While this non-random mating is the raw maternal for sexual selection in plants, whether sexual selection actually occurs and how important it may be is still nuclear.     

萝卜CMS不育系与保持系小孢子发生的细胞学研究

研究了萝卜胞质雄性不育系A2、A4及其相应保持系B2、B4的小孢子发生与花药壁发育的细胞学特征.结果表明,不育系A2的绒毡层细胞在四分体时期出现异常,小液泡增多,至单核期汇合形成大液泡,绒毡层细胞异常膨大;小孢子外壁染色浅,细胞壁受到破坏,最后与绒毡层一同降解.不育系A4在减数分裂期即表现出异常,绒毡层异常肥大;花药发育后期,小孢子外壁亦染色较浅;绒毡层细胞融合形成细胞团块侵入药室挤压小孢子,两者一同降解.     

白菜型油菜与蓝花子杂交的初步研究

通过胚胎培养,成功地获得了白菜型油菜（Ｂｒａｓｉｃａｃａｍｐｅｓｔｒｉｓ）与蓝花子（ＲａｐｈａｎｕｓｓａｔｉｖｕｓＬｖａｒｒａｐｈａｎｉｓｔｒｏｉｄｅｓＭａｋｉｎｏ）的属间杂种。该杂种具有两种类型;一种为大花类型,一种为小花类型。对它们进行花粉母细胞减数分裂的观察结果表明：小花类型为未加倍的杂种ＭＩ,存在１９个未配对染色体,大花类型为加倍或部分加倍杂种,加倍类型ＭＩ,１９个二价体排列在赤道板上;部分加倍类型ＡＩ,具有１０－１０－９的染色体组分割现象。大花类型具有可育性;它能够产生很多ｎ＝１９及ｎ＝９、ｎ＝１０的正常配子。染色体组分割能够产生倍半二倍体,它能用来研究染色体的功能和开展染色体工程。     

萝卜对土生空团菌菌丝生长的影响

纯培养条件下, 测定了十字花科植物萝卜(Raphanus sativus L.)种子、幼苗、根系分泌物及幼苗提取物对土生空团菌(Cenococcum geophilum Fr. (Cg))菌株CgSO1、CgSB2、CgO5、SPOP2 和Cg5#菌株生长的影响。结果表明供试Cg菌株与萝卜种子共培养, 或将萝卜根系分泌物和幼苗提取物加入到培养基中, 均促进了Cg菌丝生长。高温灭菌处理使萝卜根系分泌物和幼苗提取物对Cg菌株的促生作用更强, 而高温灭菌后的萝卜幼苗段对菌株生长影响不大。其中经高温灭菌处理的幼苗水提取物对5菌株的促生作用最大, CgSO1、CgSB2、CgO5、SPOP2和Cg5#每菌落的菌丝干重分别达到: 54.8、45.8、63.9、41.2和50.5 mg。     

甘蓝型油菜与蓝花子远缘杂交及双二倍体的合成研究

甘蓝型油菜品种奥罗×蓝花子杂种Ｆ-１,平均配对构型为１２．１Ⅰ＋６．５３Ⅱ＋０．４１Ⅲ＋０．１８Ⅳ＋０．１８Ⅴ,Ａ、Ｃ染色体组与Ｒ染色体间存在配对,它们之间具有一定的同源性。在甘蓝型油菜与蓝花子的杂种Ｆ-１代中,存在一种染色体不配对的减数分裂类型。这一类型中有少量可形成平衡的不减数配子。提供了油菜与蓝花子远缘杂种回交结实的细胞学根据。在ＭＳ＋０．２ｍｇ／ＬＮＡＡ＋３ｍｇ／ＬＢＡ＋１ｇ／Ｌ秋水仙碱＋３０ｇ／Ｌ蔗糖＋８ｇ／Ｌ琼脂的培养基中,接种甘蓝型油菜奥罗与蓝花子的杂种Ｆ-１进行加倍处理,经快速繁殖后,获得大量的染色体数为２ｎ＝５６的双二倍体幼苗。上述双二倍体自交结实,在减数分裂中绝大多数细胞形成２８个二价体,个别形成２６个二价体和１个四价体。上述技术在油菜与蓝花子远缘杂交中首次解决了用常规方法不易获取远种杂种稳定双二倍体的难题。甘蓝型油菜品种Ａｌｔｅｘ×蓝花子杂种Ｆ-１代,长时间的快速繁殖后,出现了染色体丢失和加倍,有形成１９条染色体的配子回复到甘蓝型油菜染色体组成的趋势。在油菜远缘杂种中发现了类似于球茎大麦远缘杂种中染色体丢失的现象。     

Effects of uniconazole and GA3 on cold-induced stem elongation and flowering of Raphanus sativus L.

The effects of gibberellin (GA) on cold-induced stem elongation and flowering of Japanese radish (Raphanus sativus L.) were investigated using application of GA3 and a GA-biosynthesis inhibitor, uniconazole (UCZ). UCZ very strongly inhibited stem elongation and delayed flowering, and the inhibition and delay were completely reversed by GA3 application. These results suggest that GA is necessary not only in the stem elongation but also in the flowering. When cold treatment (CT) was conducted on the plants whose GA level was lowered by UCZ, GA3 applied after CT completely reversed the delay of flowering. Thus low GA level probably did not retard cold induction. Microscopic observation of apical meristem showed that UCZ delayed flowering by delaying the shift from vegetative to dome-shaped meristem. This result suggested that low GA level delayed floral evocation. Consequently it was suggested that low GA level retarded physiological process involved in long day induction or in floral evocation, resulting in delay of floral evocation.     

萝卜块根中两个具溶菌酶活性的几丁质结合蛋白的纯化及其特性

通过亲和层析和羧甲基一纤维素离子交换层析从萝卜的块根中分离到两个具溶菌酶活性的酶组份：CBP1和CBP2。两者经SDS-PAGE均显示单一蛋白染色条带,其对应的分子量分别为26．9kD和24．8kD。两种蛋白除有溶菌酶活性外,还有几丁质酶活性,但无壳聚糖酶活性。各种类型的几丁质对CBP1和CBP2都有较强的吸附作用,而在还原／非还原的单向SDS-PAGE中却观察不到两者分子中存在二硫键。     

Ogura细胞质雄性不育紫菜苔×萝卜属间杂种F_1的获得及细胞遗传学研究

以Ogura细胞质雄性不育紫菜苔(AA, 2n＝20)为母本,以不同萝卜品种(RR,2n＝18)为父本进行杂交,获得了大量的属间杂种.杂种F1幼苗在低温下子叶及真叶均不缺绿.以红萝卜为父本获得的杂种F1植株叶柄、叶脉呈紫红色;以白萝卜为父本获得的杂种F1植株叶柄、叶脉不呈紫红色.所有的杂种F1植株都开白花,蜜腺正常,雄配子高度不育,但是雌配子具有部分育性.杂种F1花粉母细胞的染色体数目为预期的2n＝19,染色体平均配对构型为15.53+1.34+0.25+0.01,多数染色体以单价体的形式存在,但也有一些二价体、三价体甚至四价体,最多达到6+3+1,参加配对的染色体数达到13条,表明A染色体组和R染色体组具有一定的同源性.对该杂种的遗传及育种意义进行了讨论.     

胭脂萝卜红色素的研制

本文报道了胭脂萝卜红色素的生产工艺,并结合生产方法与相关条件开展讨论。同时报道了胭脂萝卜红色素的三种物理性状,即溶解性、坚牢度、染着性     

不同pH的酚酸溶液对杉木及萝卜幼苗生长的影响

通过四因素五水平的二次回归正交旋转试验设计 ,研究 pH值、肉桂酸、对羟基苯甲酸、香草醛四个因素对杉木和萝卜幼苗生长的影响。结果表明 ,杉木幼苗的鲜重生长随酚酸溶液的pH值的增大而增加 ,而随肉桂酸、对羟基苯甲酸和香草醛浓度的增大而减小。比较 3种不同酚类物质对杉木幼苗生长的影响 ,肉桂酸对杉木幼苗生长的影响 >对羟基苯甲酸 >香草醛 ,且 pH值与肉桂酸之间 ;肉桂酸与香草醛之间 ;对羟基苯甲酸与香草醛之间的交互作用都达到显著水平。萝卜幼苗生长对不同pH的酚酸溶液的反应与杉木幼苗相类似 ,随酚酸溶液的pH值的增大而增加 ,而随肉桂酸、对羟基苯甲酸和香草醛浓度的增大而减小。但对羟基苯甲酸对萝卜幼苗生长的影响 >肉桂酸 >香草醛 ,酚酸溶液的 pH值与所有 3种酚酸之间的交互作用都达到显著水平 ;肉桂酸与香草醛的交互作用对萝卜幼苗生长的影响也达到F0 0 1水平显著。     

LED不同光质对萝卜愈伤组织诱导、增殖和萝卜硫素含量的影响

以白水萝卜无菌苗及其愈伤组织为实验材料,研究其在LED白、红、黄、蓝、绿和蓝红6种光质下的愈伤诱导和增殖。结果表明：LED不同光质下胚轴愈伤组织的诱导效应不同,诱导率顺序依次为黄光〉红光〉蓝红光〉白光〉蓝光〉绿光;蓝光、黄光和红光有利于子叶愈伤组织的诱导;子叶诱导愈伤组织的效果较下胚轴好;LED红光下愈伤组织的增殖倍数和萝卜硫素含量均为最高。     

Response of seedlings of radish (Raphanus sativus) to osmotic shock and external hydrostatic pressure

The effects of an osmotic shock (300 m M mannitol, –0. 67 MPa) or/and increased external hydrostatic pressure on seedlings (42-h-germinated seeds) of radish ( Raphanus sativus L. cv. Tondo Rosso Quarantino) were investigated. The osmotic shock did not inhibit H⁺ extrusion and net K^- uptake, and did not affect the levels of malic acid, reducing sugars, sucrose or amino acids or of the energy charge (i. e. the synthesis of energy-rich phosphate bonds), but inhibited the synthesis of proteins. RNA and DNA, measured as incorporation of labelled precursors. When the osmotic shock was applied together with an increased external hydrostatic pressure of the same magnitude (+0. 67 MPa), the same metabolic parameters and the inhibition of synthesis of RNA and DNA were not substantially affected, while the inhibition of protein synthesis was practically reversed and the energy charge decreased; the recovery of protein synthesis was not due to a change in labelled leucine uptake capability. Increased external hydrostatic pressure alone decreased the energy charge without affecting the other parameters considered.
The possibility that protein synthesis activity is directly controlled by cell turgor pressure (internal hydrostatic pressure) is discussed.     

Efficient Agrobacterium-mediated genetic transformation method using hypocotyl explants of radish (Raphanus sativus L.)

To investigate the gene function of radish (Raphanus sativus L.), several attempts have been made to generate genetically transformed radish. However, no efficient and relatively simple method for the genetic transformation of radish has been developed to date. In this study, we established an Agrobacterium-mediated genetic transformation method using the hypocotyl-derived explants of radish cultivar “Pirabikku”. Primarily based on the Brassica transformation procedure, we optimized it for radish transformation. Using this system, the transformation efficiency of radish hypocotyl explants by Agrobacterium tumefaciens strain GV3101 harboring pIG121-Hm was 13.3%. The copy number of transfer DNA integrated into the genome was either one or two in the four independent transgenic plants. Two of the four plants exhibited male sterility and did not produce self-pollinated seeds. Examination of the expression of the β-glucuronidase (GUS) gene in T₁ plants from fertile T₀ plants showed that the GUS genes were inherited. The improvement in the genetic transformation in this study might pave the way for accelerated molecular breeding and genetic analysis of radish.     

Accumulation of a 22-kDa protein and its mRNA in the leaves of Raphanus sativus in response to salt stress or water deficit

The response of 10-day-old seedlings of Raphanus sativus L. cv. Fakir to salt stress (100 m M to 200 m M NaCl) was investigated. Three weeks after initiation of salt treatment, the fresh weight of the shoots of salt-treated plants was half that of untreated plants. The salt stress resulted in the accumulation of Na⁺, preferably in the old leaves. The K⁺ level was reduced by as much as 50% in the old leaves of NaCl-treated plants, whereas this reduction was only 20–25% in the young leaves. Free proline accumulated in all aerial organs, and the highest levels were found in the young leaves. Patterns of total proteins extracted from the leaves of control or salt-treated plants were compared. The most obvious change concerned a 22-kDa, pl 7.5 polypeptide, which accumulated after exposure of the plants to NaCl. The appearance of this polypeptide was also mediated by a rapid drought stress, and sequencing indicated that it is related to the Künitz protease inhibitor family. A cDNA clone corresponding to the radish 22-kDa polypeptide was obtained and sequenced. Northern blot analysis showed that salt stress induces a large accumulation of this mRNA in the leaves of radish.