摘要
试材'98-2'(Vitis vinifera cv.98-2)和‘洛浦早生’(V.vinifera cv. Luopuzaosheng)为河南科技大学园艺系分别从‘巨峰’(V.vinifera cv. Kyoho)和‘京亚’(V.vinifera cv. Jingya)中选育的早熟葡萄芽变新品种(系)。本研究对‘98-2’和‘洛浦早生’与其亲本的植物学和生物学特性及果实成熟过程中矿质元素、激素、糖、酸的变化以及分子鉴定等方面进行了研究,探讨了芽变品种(系)与亲本上述特性的差异,取得以下主要结果：
1.通过对芽变品种(系)与其亲本的诸多植物学性状和生物学特性指标的比较分析,发现两个芽变品种(系)在果实大小、可溶性固形物含量、可滴定酸含量、物候期、果穗及果粒呼吸强度和乙烯释放量等方面均有不同程度的差异,以果实成熟期差异较为明显。果实发育期,‘98-2’比‘巨峰’少36 d左右,‘洛浦早生’比‘京亚’少20 d左右。
2.果实发育期间矿质元素变化研究表明,在幼果期果实中除Ca外,其它元素的含量均是‘巨峰’高于‘98-2’；‘洛浦早生’除Fe、Zn、N、P、K等矿质元素外,其余均高于‘京亚’。此后,随着幼果的快速增大,因稀释作用,使多数矿质元素含量逐渐下降。进入果实迅速膨大期后,只有Mn元素出现一个较大值,其它大多数矿质元素平稳递减。在果实成熟时,‘98-2’果实中Zn、Ca、Mg、P、K、元素的含量高于‘巨峰’,其余则为巨峰高于‘98-2’；而‘洛浦早生’除Cu、Zn、Mg、P元素外,其余均是‘京亚’为高。
3.果实发育期果肉中IAA、GA3、ZR、ABA的含量变化趋势相似,都是一个单峰曲线,在果实缓慢生长期开始前后即花后30 d或40 d达到较大值,但较大值出现时期与成熟期早晚无关。果皮中IAA、GA3、ZR的含量变化趋势相似,都有一明显的较大值,两个芽变品种(系)的较大值均低于亲本,且较大值出现时间可能与成熟期早晚有关；果皮中ABA含量‘98-2’高于‘巨峰’,‘洛浦早生’其含量低于‘京亚’,果皮中ABA含量在果实发育的后期含量较高,但成熟期不同的品种高含量ABA所持续的时间不同。
4.果实发育期除蔗糖外,果肉中果糖、葡萄糖及总糖的含量变化‘98-2’与‘巨峰’有差别,‘洛浦早生’和‘京亚’相似。‘巨峰’果肉中果糖、葡萄糖及总糖的含量在花后40 d前增加缓慢,以后则迅速增加直到果实成熟：‘98-2’和‘京亚’分别在花后50 d、‘洛浦早生’在花后40 d以后果肉中果糖、葡萄糖及总糖的含量开始降低直到果实成熟,但‘洛浦早生’果实发育后期果糖、葡萄糖及总糖的含量均高于‘京亚’
果皮中果糖、葡萄糖及总糖的含量变化‘98-2’、‘巨峰’、‘洛浦早生’、‘京亚’相似。花后30 d内4个品种果皮中果糖、葡萄糖及总糖的含量变化不大,以后则逐渐上升直到果实成熟,且在果实成熟时芽变品种(系)果皮中果糖、葡萄糖及总糖的含量均低于亲本。
果实发育期转化酶均在果实糖分开始快速积累前后活性达到最大,‘洛浦早生’与‘京亚’变化较一致,而‘98-2’与‘巨峰’差别较大。
5.无论芽变品种(系)和亲本,果实中酸的含量均以酒石酸、苹果酸最多,柠檬酸次之,草酸最低。果肉中4个品种苹果酸含量随果实生长发育逐渐上升,在果实发育中期上升到较大值,成熟期则下降,且芽变品种(系)的苹果酸含量均低于亲本；而果皮中苹果酸含量只有‘98-2’前期高于亲本,其余则和果肉的变化趋势一样。果肉、果皮中的酒石酸含量,在生长至成熟过程中均呈下降趋势,且芽变品种(系)酒石酸含量均低于亲本。果肉中总酸含量与苹果酸变化趋势相似,果皮中则与酒石酸相似。
6.利用RAPD分子标记技术对两个芽变品种(系)与亲本的DNA差异进行了分析,发现S56的PCR产物中条带1300 bp在‘98-2’中出现,而‘巨峰’中没有出现,S234的PCR产物中条带950 bp只在‘京亚’中出现,而‘洛浦早生’中出现条带1320 bp。利用SSR标记,引物VRZAG25和VVS2分别在巨峰及其芽变98-2,京亚及其芽变洛浦早生的PCR产物中出现510bp和290bp的差异条带。从分子证据上表明它们是不同的品种,证明了它们具有芽变关系的可能。
7.‘98-2’、‘巨峰’、‘京亚’与‘洛浦早生’的花粉粒形态相似；只是‘98-2’较细长,‘巨峰’较粗短；‘京亚’与‘98-2’、‘洛浦早生’的花粉形状也略有不同。花粉的萌发率‘98-2’低于‘巨峰’,‘京亚’高于‘洛浦早生’。
8.‘98-2’与‘巨峰’的POD同工酶谱存在差异,‘98-2’与‘巨峰’相比多了一条P3带而少了一条P6带,而且P4、P5为强带。表明‘98-2’和‘巨峰’不是同一种种质,它们的遗传物质不完全相同。而‘洛浦早生’和‘京亚’的同工酶谱完全相同。
Two new grape varieties,'98-2'(Vitis vinifera) and 'Luopuzaosheng'(V. viniferd) were early-ripening bud mutations varieties of 'Kyoko'(V. vinifera) and 'Jingya'(V. vinifera), respectively. Both of them were selected by college of Horticulture, Henan University of Science & Technology. In this study, the botanical and biological characteristics, the variations of mineral elements, hormone, sugar and acid, as well as molecular identification were studied among the varieties ('98-2' and 'Luopuzaosheng') and their parents. The difference between the bud mutation varieties and their parents were also discussed. The results were as follows:
1. There are some differences in botanical characters and biological characteristics between the bud mutation varieties (strain)and their maternal plants, such as the fruit size, the content of soluble solid and titratable acid, the phenophase, the respiration rate of fruit cluster and fruit grain, the ethylene release. Moreover, the most significant variation was fruit ripening period. The fruit developing period of the '98-2' was 36d shorter than 'Kyoko', while the 'Luopuzaosheng' was 20d shorter than Jingya.
2. Theanalyis of the content variations of mineral elements of fruit in fruit developing period showed the content of other mineral elements in young fruits of 'Kyoko' was higher than '98-2' except for Ca. In Jingya's young fruit the content of Fe, Zn, N, P and K was higher than 'Luopuzaosheng', but other mineral elements were less than 'Luopuzaosheng'. After entering the fruit quickly expanding period, the content of the majority of mineral elements gradually decreased except for Mn due to the diluting effect. When the fruit matured, the content of Zn, Ca, Mg, P and K in '98-2' was higher than 'Kyoko' while other mineral elements was lower than 'Kyoko'. The content of Cu, Zn, Mg and P in 'Luopuzaosheng' was higher than 'Jingya' and the other mineral elements were lower than 'Jingya'.
3. At the fruit developing period, the change trends of the content of IAA, GA3, ZR and ABA in flesh were similar which appear as a single peak profile. The content of IAA, GA3, ZR and ABA present the peak in 30 or 40 days after flowering at the beginning of the fruit slowly growth period. But the time of the peak value emerging has no relationship with the maturity period. The change trends of the content of IAA, GA3 and ZR in peel were similar which all have a significant peak. The peak values of the two bud mutation varieties (strain) were lower than their parents and the time of the peak valueemerging have relationship with the maturity period. The content of ABA in peel of '98-2' was higher than 'Kyoko', but 'Luopuzaosheng' was lower than 'Jingya'. Moreover the content of ABA was very high in the later stage of fruit developing period, but the continuous time of high content of ABA was different in different maturity period cultivars.
4. At the fruit developing period, the change trends of the content of frutose, glucose and total sugar in flesh were different between '98-2' and 'Kyoko', but they were similar among 'Luopuzaosheng' and 'Jiangya'. The content of frutose, glucose and total sugar increased slowly during forty days after flowering and then increased rapidly until the fruit matutity. The content of frutose, glucose and total sugar began to decrease at 50d after flowering of'98-2'and'Jingya' and at 40d after flowering of 'Luopuzaosheng'. For 'Luopuzaosheng', the content of frutose, glucose and total sugar was higher than 'Jingya' in the later stage of fruit developing period.
The variation of the content of frutose, glucose and total sugar were similar among the '98-2','Kyoko','Luopuzaosheng' and 'Jingya' in peel at the period of the fruit developing. During thirty days after flowering, the content of frutose, glucose and total sugar had little changes in peel of these four varieties(strain). Then they rose gradually till the fruit maturity. Furthermore the content of frutose, glucose and total sugar in peel of bud mutation were lower than their parents'
The activity of the invertase reached the maximum at the beginning of the fruit sugar rapidly accumulating in the fruit developing period. This variations were similar between 'Luopuzaosheng' and 'Jingya', but there are great difference between '98-2' and 'Kyoko'.
5. The content of the tartaric acid and malic acid was the most abundant in the fruit and the next was citric acid, the lowest was oxalic acid whether in bud mutation varieties(strain) or parents. The content of the malic acid was gradually increased with the fruit developing in the flesh of these four varieties(strain). Then it reached to the highest value in the middle period of berry fruit developing, and decreased in maturity. Meanwhile the content of the malic acid of bud mutation was lower than parents. In peel the content of the malic acid of '98-2' was higher than parents' at earlier stage and the change trend was the same as it in flesh in other phase. The contents of tartaric acid all showed downtrend whether in flesh or in peel from growth to maturity. Furthermore the contents of tartaric acid of bud mutation were lower than parents. The trend of content of total acid in flesh was similar to the trend of malic acid but in peel it was similar to the trend of tartaric acid.
6. There are difference at DNA level between the two bud mutation varieties(strain) and their parents by using RAPD and SSR markers. The results showed there was a band (1300bp) amplied by RAPD prime S56 in 98-2 but not in Kyoho. A band (950bp) amplied by RAPD primer S234 was appeared only in Jingya but a band (1320bp) amplied by the same primer was appeared in Luopuzaosheng. SSR primers, VRZAG25 and VVS2 also amplified two distinct bands,510bp and 290bp, in each two varieties with the relationship of bud mutation (Kyoho and 98-2, Luopuzaosheng and Jingya). These results proved that they are not the same varieties and probably have the bud mutation relationship.
7. The pollen shapes of '98-2','Kyoho','Jingya' and 'Luopuzaosheng' were similar. The '98-2" s pollen grain was spindly and 'Kyoho" s was dumpy and 'Jingya" s had slightly different with '98-2" s and 'Luopuzaosheng" s. Pollen germination percentage of '98-2' was lower than that of 'Kyoho' and 'Jingya' was higher than 'Luopuzaosheng'.
8. Comparizing with the band patterns of peroxidase isoemzyme between '98-2' and 'Kyoho','98-2' had a P3 band but P6 band was absent and the P4 and P5 band were strong bands which illustrated that their genetic materials were not identical. The band patterns of isoemzyme between 'Luopuzaosheng' and 'Jingya' were completely identical.
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