节点文献
野生鸭儿芹组织培养及其种子休眠特性、破除方法的研究
Tissue Culture, Dormancy Break Approaches and Property of Dormant Seeds of Wild Cryphtotaenia Japonica
【作者】 喻梅;
【导师】 周守标;
【作者基本信息】 安徽师范大学 , 植物学, 2011, 硕士
【摘要】 鸭儿芹(Cryptotaenia japonica Hassk.),又名鸭脚板,属于鸭儿芹属(Cryptotaenia DC.)伞形科(Umbelliferae)植物,为药食两用植物,嫩苗及叶可供蔬食,全草及根可提取芳香油,全草及果可入药,果实可榨油,供制肥皂和油漆用。目前国内外学者对其研究还不多,主要集中在对其营养成分及矿质元素的分析、食品的加工研制、挥发性成分研究、药用及毒理研究、形态解剖学研究及栽培技术研究等方面。以安徽地区的野生鸭儿芹为研究材料,以鸭儿芹叶作为外植体对野生鸭儿芹的组织培养进行了初步研究;以野生鸭儿芹种子为实验材料,研究了鸭儿芹种子的休眠特性和解除方法,并对安徽四地区的鸭儿芹种子萌发特性进行了研究,为安徽种植鸭儿芹的选种提供依据和基础资料。结果如下:1.研究了鸭儿芹不同材料(茎、叶)作为外植体诱导愈伤组织的最佳条件,愈伤组织继代、试管苗生根及试管苗移栽所需的条件。结果表明固体培养基为MS+6-BA 2.0 mg/L + NAA 1.0 mg/L对叶片诱导愈伤组织较好,诱导生根的最佳培养基为1/2MS + IAA 1.0 mg/L,较适合鸭儿芹愈伤组织继代的培养基为MS+6-BA 1.0 mg/L + NAA 0.1 mg/L,在试管苗的移栽中,选用珍珠岩∶蛭石∶腐殖土=1∶1∶1的基质移栽的成活率较腐殖土基质效果更好。2.TTC法对种子活力的测定表明有活力的种子为55.33%±3.71%;切破种皮种子与完整种子吸水率在前12 h相差较大,但最终吸水率相差不大,分别达到70.00%±1%和68.32%±0.32%,表明种皮并不阻碍种子吸水;种子中存在内源抑制物,其粗提液在较低浓度下即可抑制芹菜种子的萌发;鸭儿芹种子成熟时胚未分化完全,胚率为28.65%±2.49%,经过低温处理后完成后熟,胚率达到65.93%±3.86%,萌发率达到100.00%,因此鸭儿芹种子具有形态生理休眠特性。清水浸种和低温冷藏共同处理可有效解除其休眠,浸种和低温冷藏具有交互效应,浸种36 h、5°C冷藏30天即可解除其休眠,萌发率达到100.00%,发芽势达到91.11%±0.91%。已破除休眠的种子适宜其萌发的温度范围扩大(15.0~27.5°C ),而且在土壤中也可较好的萌发,萌发率达到96.67%±3.33%,发芽势达到71.11%±1.93%。3.安徽四产地(黄山、大别山、繁昌、南陵)的鸭儿芹种子千粒重、外部形态(种子长、宽、厚)有差异,千粒重:黄山( (2.67±0.15)g)>大别山( (1.98±0.06)g)>繁昌( (1.88±0.06)g)>南陵( (1.57±0.01)g);休眠解除前,种子活力、相对电导率值不同,种子活力:大别山(78.67%±1.76%)>繁昌(74.00%±3.06%)>黄山(64.67%±4.67%)>南陵(55.33%±3.71%),相对电导率测定结果:大别山(23.20%±2.23%)<繁昌(23.70%±1.00%)<黄山(35.91%±2.79%)<南陵(43.54%±4.04%);破除休眠后,在滤纸上的发芽势、萌发率几乎差不多,在土壤中的发芽势、萌发率以及幼苗成活率不同,四产地种子萌发率没有显著性差异,南陵地区种子发芽势和幼苗成活率显著低于其他三个产地,黄山、大别山、繁昌三产地种子的发芽势、萌发率以及幼苗成活率无显著性差异。
【Abstract】 Cryptotaenia japonica Hassk (Umbelliferae), a perennial herbaceous plant, is native to China and Japan. It usually favors humid eco-environment. This plant species is both an edible and pharmaceutical plant. C. japonica is a popular vegetable in Japan and usually served as salad or prepared in soup. But in China it is usually cooked and then served. This plant can be cultivated year round in green houses. And due to easy handling, C. japonica is also grown hydroponically. Its nutritious and pharmaceutical nature are of great concerns among agriculturalists and farmers. The aromatic ingredient in the whole plant also has commercial and industrial potential. To the best of my knowledge, only a few studies have been addressed to C. japonica as yet. And these studies have been concentrating on the nutrient and mineral elements, its potential as dietary food, the volatile constituent, the pharmaceutical purposes and toxicology, and its morphological anatomy. No studies however according to my knowledge have ever tried to investigate its ontogenesis like seed germination and seed dormancy. Our studies will lend support to the artificial cultivation of C. japonica. C. japonica seeds are characterized by a long dormancy period and will not germinate without any treatment, thereby impeding its agricultural cultivation. The objectives of the present research were to investigate the reasons for seed dormancy and optimal approaches to dormancy break of C. japonica seed and to supply theoretical fundamentals for its artificial cultivation..The paper made detailed studies on tissue culture, the dormant seeds break approaches and the germination property of C. japonica from Anhui province. It could afford some basic scientific data for the plant extensively in Anhui province.The results were as follows:1. In the tissue culture of C. japonica, the callus was induced from the stems, leaves, many experiments was made to explore the optimal condition of succesive transfer cultue, root induction of test-tube plant and its transplant. The results showed that the culture medium with MS+6-BA 2.0 mg/L + NAA 1.0 mg/L is suitable for the inducling callus from leaves, the culture medium with 1/2MS+IAA 1.0 mg/L is suitable for the induction of adventitious root, the culture medium with MS+6-BA 1.0 mg/L + NAA 0.1 mg/L is suitable for succesive transfer cultue, and in the transplant of the test-tube plant it was very effective with the substance: perlite : vermiculite : humus =1 : 1 : 1, the percentage of the survival seeding were higher.2. The results showed that, based on TTC (triphenyltetrazolium chloride) staining method, only 55.33%±3.71% of the seeds were viable. Although the imbibition rates of broken and intact seeds differ markedly in the early 12 h, the final imbibition rates were almost the same, 70.00%±1% and 68.32%±0.32%, respectively, suggesting that seed imbibition is not impeded by seed coat. The seed germination of celery was inhibited by the low-concentration crude extracts of C. japonica because there was probably endogenous inhibitants. The embryo of matural C. japonica seed was not developed completely since the percentage of the length of embryo/length of endosperm was 28.65%±2.488%. After post maturation under low temperature it was close to 65.93%±3.86% and the germination rate was almost 100.00%. Therefore the seed of C. japonica was featured by morphophysiological dormancy. Both soaking with tap water and storage under low temperature can break effectively the seed dormancy of C. japonica and both methods interact. Soaking for 36 h and storage at 5°C for 30d can break seed dormancy and the germination rate and germination potential can be close to 100.00% and 91.11±0.91%, respectively. The optimal temperature for the germination of dormany-broken seeds was 15.0~27.5°C and better germination rate and germination potential in soil can be 96.67%±3.33% and 71.11%±1.93%, respectively.3. There was significance difference on thousand kernels weight and seed shape (length, width, thickness) of four areas in Anhui province. The result of thousand kernels weight showed that: Huangshan ( (2.67±0.15)g) > Dabieshan ( (1.98±0.06)g) > Fanchang ( (1.88±0.06)g) > Nanling ( (1.57±0.01)g); The seed vitality and the relative electric conductivity are significance difference before the dormany-broken, the result of the seed vitality is follows: Dabieshan (78.67%±1.76%) > Fanchang (74.00%±3.06%) > Huangshan (64.67%±4.67%) > Nanling (55.33%±3.71%); the result of the relative electric conductivity is follows : Dabieshan (23.20%±2.23%) < Fanchang (23.70%±1.00%) < Huangshan (35.91%±2.79%) < Nanling (43.54%±4.04%); After overcome the dormancy, there was no difference about the germination energy and germination rate on the filter paper, but when in the soil the ermination energy energy and the survival seeding rate were significance differences, the ermination energy energy and the survial seeding rate of the Nanling seeds were low than other three areas. There was no difference on germination energy, germination rate and the survival seeding rate of Huangshan, Dabieshan and Fanchang those three areas .
【Key words】 Cryphtotaenia japonica Hassk; tissue culture; seeds; morphophysiological dormancy; germination rate; areas;