CN115266617A - A screening method for low temperature-tolerant garden chrysanthemum varieties in plateau areas - Google Patents
A screening method for low temperature-tolerant garden chrysanthemum varieties in plateau areas Download PDFInfo
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- 238000005520 cutting process Methods 0.000 claims description 43
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 claims description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 26
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- NSMUHPMZFPKNMZ-VBYMZDBQSA-M chlorophyll b Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C=O)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 NSMUHPMZFPKNMZ-VBYMZDBQSA-M 0.000 claims description 9
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Abstract
Description
技术领域technical field
本发明涉及一种园林小菊的筛选方法，尤其涉及一种高原地区耐低温园林小菊品种的筛选方法，属于菊花品种筛选技术领域。The invention relates to a screening method for small garden chrysanthemums, in particular to a screening method for low-temperature-resistant garden small chrysanthemum varieties in plateau areas, and belongs to the technical field of chrysanthemum variety screening.
背景技术Background technique
菊花(Chrysanthemum morifolium)为菊科菊属多年生宿根草本植物，其中园林小菊具有株型矮小、紧凑、品种繁多、观赏性强的特点成为了一种城市绿化的良好地被材料，在生产中采用无性的扦插繁殖，因其生长快、繁殖系数高，使其在花卉市场中占有十分重要的地位。Chrysanthemum (Chrysanthemum morifolium) is a perennial herbaceous plant of the genus Chrysanthemum in the family Asteraceae. Among them, the garden chrysanthemum has the characteristics of short plant size, compactness, variety, and strong ornamental characteristics. It has become a good ground cover material for urban greening. Due to its fast growth and high reproduction coefficient, it plays an important role in the flower market.
近年来，随着环境建设和新品种选育，各个地区都引进了很多菊花品种，但菊花再生长过程中对温度的要求比较严格，温度作为影响扦插的关键因素之一，过高或过低都不利于新根萌发与生长，甚至低温会导致菊花扦插苗出现生根慢、根系和插穗发育不良等现象(Tsipouridis et al.,2005；李晓伟等，2014)。In recent years, with the construction of the environment and the selection of new varieties, many chrysanthemum varieties have been introduced in various regions, but the temperature requirements for the regrowth of chrysanthemums are relatively strict. As one of the key factors affecting cuttings, temperature is too high or too low. Both are not conducive to the germination and growth of new roots, and even low temperature will lead to slow rooting of chrysanthemum cuttings, poor development of roots and cuttings (Tsipouridis et al., 2005; Li Xiaowei et al., 2014).
发明人所处的青海地区也引进了很多菊花品种。但因青海地处青藏高原，海拔高、气温低，严重影响着扦插苗的生长发育，极大地降低了扦插育苗的成活率，导致扦插苗生根时间长、根系质量差以及插穗生长发育不良等问题。虽然大部分生产已经采用日光温室，但由于总体基础薄弱，其光照、温度和可控性较差，加之加温系统育苗成本高，春季低温已成为高原地区菊花设施栽培的制约因素，造成插穗生根所需的时间长于其他地区。因此，青海地区引进菊花品种时多引进生根苗，其价格高、体积大严重影响了菊花在青海地区的应用。有关园林小菊耐低温品种筛选及适应性评价的研究还未见报道，进一步造成了青海地区菊花品种引进时的盲目性。The Qinghai area where the inventor is located has also introduced a lot of chrysanthemum varieties. However, because Qinghai is located on the Qinghai-Tibet Plateau, the altitude is high and the temperature is low, which seriously affects the growth and development of cutting seedlings, greatly reducing the survival rate of cutting seedlings, resulting in long rooting time of cutting seedlings, poor root quality and poor growth and development of cuttings. . Although solar greenhouses have been used for most of the production, due to the weak overall foundation, poor light, temperature and controllability, coupled with the high cost of raising seedlings in the heating system, the low temperature in spring has become a restrictive factor for the protected cultivation of chrysanthemums in plateau areas, resulting in rooting of cuttings The time required is longer than other regions. Therefore, when introducing chrysanthemum varieties in Qinghai area, rooted seedlings are often introduced, and their high price and large size have seriously affected the application of chrysanthemums in Qinghai area. The research on the screening and adaptability evaluation of low-temperature tolerant varieties of garden chrysanthemum has not been reported, which further caused the blindness of the introduction of chrysanthemum varieties in Qinghai area.
发明内容Contents of the invention
针对现有技术的上述缺陷，本发明旨在提供一种简单而又符合实际生产所需的园林小菊耐低温品种的筛选方法。Aiming at the above-mentioned defects of the prior art, the present invention aims to provide a screening method for low-temperature-resistant garden Chrysanthemum species that is simple and meets the needs of actual production.
本发明所述的一种高原地区耐低温园林小菊品种的筛选方法，包含以下步骤：A kind of screening method of low-temperature-resistant garden small chrysanthemum varieties in the plateau area of the present invention comprises the following steps:
(1)剪取园林小菊插穗，扦插于穴盘中，设2个温度进行扦插处理：胁迫组昼/夜15℃/15℃和对照组25℃/20℃；(1) Cut the cuttings of garden chrysanthemum, put the cuttings in the plug tray, and set 2 temperatures for cutting treatment: the stress group day/night 15°C/15°C and the control group 25°C/20°C;
(2)测定园林小菊插穗根总表面积和根尖数：在园林小菊生长期，测定其根总表面积和根尖数，分别以cm2和根为单位；(2) Determination of the total surface area of cuttings and the number of root tips of Chrysanthemum chrysanthemum cuttings: in the growth period of Chrysanthemum chrysanthemum, measure the total surface area of its roots and the number of root tips, taking cm and root as unit respectively ;
(3)测定园林小菊插穗总叶绿素含量：取待测园林小菊鲜样用乙醇提取叶绿素，将叶绿素提取液倒入比色皿中，以乙醇溶液作为对照并进行调零，测定波长665nm和649nm下的吸光度；按公式计算出叶绿素a、叶绿素b和总叶绿素浓度，以获得组织中总叶绿素含量；其中：叶绿素a浓度＝13.95A665-6.88A649；叶绿素b浓度＝24.96A649-7.32A665；总叶绿素浓度＝叶绿素a浓度+叶绿素b浓度；叶绿体色素的含量＝(色素浓度*提取液体积)/样品鲜重；(3) Determination of the total chlorophyll content in cuttings of Chrysanthemum chrysanthemum: take fresh samples of Chrysanthemum chrysanthemum to be tested and extract chlorophyll with ethanol, pour the chlorophyll extract into a cuvette, use ethanol solution as a control and perform zero adjustment, and measure at wavelengths of 665nm and 649nm Absorbance; calculate chlorophyll a, chlorophyll b and total chlorophyll concentration according to the formula to obtain the total chlorophyll content in the tissue; wherein: chlorophyll a concentration=13.95A 665-6.88A 649 ; chlorophyll b concentration= 24.96A 649-7.32A 665 ; Total chlorophyll concentration=chlorophyll a concentration+chlorophyll b concentration; Chloroplast pigment content=(pigment concentration*extract volume)/sample fresh weight;
(4)计算步骤(2)和步骤(3)测定的指标的耐低温系数：根总表面积耐低温系数X1＝(胁迫后测定值/对照测定值)*100％；总叶绿素含量耐低温系数X2＝(胁迫后测定值/对照测定值)*100％；根尖数耐低温系数X3＝(胁迫后测定值/对照测定值)*100％；(4) Calculate the low temperature resistance coefficient of the index measured in step (2) and step (3): root total surface area low temperature resistance coefficient X 1 =(measured value after stress/contrast measured value)*100%; total chlorophyll content low temperature resistance coefficient X 2 =(measured value after stress/measured value of control)*100%; low temperature tolerance coefficient of root tip number X 3 =(measured value after stress/measured value of control)*100%;
(5)计算耐低温性综合评价值＝0.220+0.090X1+0.580X2+0.095X3；耐低温性综合评价值评分越高，且X1、X2和X3均不为零时，为越耐低温品种。(5) Calculate the comprehensive evaluation value of low temperature resistance = 0.220+0.090X 1 +0.580X 2 +0.095X 3 ; the higher the comprehensive evaluation value of low temperature resistance, and X 1 , X 2 and X 3 are not zero, For the more low temperature tolerant varieties.
本发明步骤(1)剪取园林小菊的插穗可以按照本领域的常规方法，在一种具体的实例中，剪取园林小菊8～10cm的顶梢为插穗，留上部2～3片叶，扦插于穴盘中。Step (1) of the present invention cuts the cuttings of garden chrysanthemum according to the conventional methods in the art. In a specific example, the cuttings of 8-10 cm of garden chrysanthemum are cut as cuttings, and the top 2-3 leaves are left for cutting. in the hole tray.
本发明步骤(2)的园林小菊生长期是指扦插后26～28天。The growth period of small garden chrysanthemum in step (2) of the present invention refers to 26-28 days after cutting.
本发明步骤(2)测定园林小菊插穗根总表面积和根尖数可以按照本领域常用的根系扫描系统，例如根系表型分析系统GXY-A、根系表型分析软件Win-RHIZO等根系扫描分析软件。Step (2) of the present invention measures the root total surface area of cuttings of Chrysanthemum chrysanthemum cuttings and the number of root tips according to the root scan system commonly used in the art, such as root scan analysis software such as root system phenotype analysis system GXY-A, root system phenotype analysis software Win-RHIZO, etc. .
本发明步骤(3)测定园林小菊插穗总叶绿素含量可以按照本领域的常规方法，为了使测定结果更具统一和准确性，本申请提供一种具体的测定方法：分别取胁迫组和对照组的待测园林小菊鲜样，洗净表面污物，剪碎混匀，各称取剪碎鲜样0.1g，共3份，分别置于10mL离心管中；每个离心管中加入5mL 95％的乙醇，使样品完全浸泡在乙醇溶液中；置于暗处浸泡24h后，将叶绿素提取液倒入比色皿中，以95％乙醇溶液作为对照并进行调零，测定波长665nm和649nm下的吸光度。The step (3) of the present invention measures the total chlorophyll content of garden Chrysanthemum cuttings according to conventional methods in the art. In order to make the measurement results more uniform and accurate, the application provides a specific measurement method: take the stress group and the control group respectively. For fresh garden chrysanthemum samples to be tested, wash the surface dirt, chop and mix well, weigh 0.1g of each fresh sample, and put them into 10mL centrifuge tubes respectively; add 5mL of 95% Ethanol, so that the sample is completely immersed in the ethanol solution; after soaking in a dark place for 24 hours, pour the chlorophyll extract into a cuvette, use 95% ethanol solution as a control and perform zero adjustment, and measure the absorbance at wavelengths 665nm and 649nm .
本发明步骤(5)耐低温性综合评价值评分越高，为越耐低温品种，通常当评分≥0.85时，为耐低温园林小菊品种，即在10℃～25℃的温度下扦插均能够生长的品种。In step (5) of the present invention, the higher the score of the low temperature resistance comprehensive evaluation value is, the more low temperature resistant variety is. Usually when the score is ≥0.85, it is a low temperature resistant garden chrysanthemum variety, that is, the cuttings can grow at a temperature of 10°C to 25°C. varieties.
在一些实施例中，本发明所述的园林小菊具体为金陵粉玉兰、金陵红荷、金陵礼花、金陵笑靥、金陵一品红或金陵潋滟。In some embodiments, the garden chrysanthemum described in the present invention is specifically Jinling pink magnolia, Jinling red lotus, Jinling firework, Jinling smile, Jinling poinsettia or Jinling twinkle.
本发明相对于现有技术的有益效果：The beneficial effect of the present invention relative to prior art:
本发明针对高原地区园林小菊耐低温品种的筛选提供了一种简便有效的方法，通过园林小菊2个根系指标和1个光合色素指标的测定，可进行园林小菊低温适应性评价，从而筛选园林小菊的耐低温品种。本发明所述的方法只需通过测定3个指标，就能进行园林小菊耐低温品种的鉴定，不仅鉴定成本低，而且操作简单，十分易于推广。The present invention provides a simple and effective method for the screening of small garden chrysanthemum low-temperature-resistant varieties in plateau areas. Through the determination of two root system indexes and one photosynthetic pigment index of small garden chrysanthemum, the low temperature adaptability of small garden chrysanthemum can be evaluated, thereby screening small garden chrysanthemum low temperature tolerant varieties. The method of the invention can identify low-temperature-resistant varieties of small garden chrysanthemums only by measuring three indicators, and not only has low identification cost, but also is simple to operate and is very easy to popularize.
本发明所述的方法，建立了较可靠的综合评价方法，能够有效的筛选出耐扦插期低温品种，为高原地区园林小菊引进和产业具有重要意义。The method of the invention establishes a relatively reliable comprehensive evaluation method, can effectively screen out low-temperature-resistant varieties in the cutting period, and is of great significance for the introduction and industry of small garden chrysanthemums in plateau areas.
具体实施方式Detailed ways
以下的实施例便于更好地理解本发明，但并不限定本发明。下述实施例中的实验方法，如无特殊说明，均为常规方法。下述实施例中所用的试验材料，如无特殊说明，均为自常规生化试剂商店购买得到的。The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores.
若无特别说明，以下实施例的实验材料如下：由南京农业大学引进的6个园林小菊品种作为试材，分别为金陵潋滟(粉紫)、金陵笑靥(紫色)、金陵一品红(红色)、金陵礼花(复色)、金陵粉玉兰(粉紫色)、金陵红荷(红色)。花期为8月中旬至9月下旬与青海的旅游季有交集。6个园林小菊品种扦插苗定植于青海大学园艺所试验地大棚内，采用随机区组，株行距为40×40cm，并且采取相同的常规水肥管理方法。Unless otherwise specified, the experimental materials of the following examples are as follows: 6 garden chrysanthemum varieties introduced by Nanjing Agricultural University are used as test materials, namely Jinling Lianyan (pink purple), Jinling Xiaodi (purple), Jinling poinsettia (red), Jinling Fireworks (recolor), Jinling Pink Magnolia (pink purple), Jinling Honghe (red). The flowering period is from mid-August to late September, which coincides with Qinghai's tourist season. The cutting seedlings of 6 garden Chrysanthemum varieties were planted in the greenhouse of the experimental field of the Horticultural Institute of Qinghai University, using random blocks, the spacing between plants and rows was 40×40cm, and the same conventional water and fertilizer management methods were adopted.
实施例1Example 1
本发明对引进的6个南京农业大学的园林小菊品种进行耐低温品种筛选，具体供试材料如表1所示。The present invention screens low-temperature-resistant varieties of 6 imported garden chrysanthemum varieties from Nanjing Agricultural University, and the specific materials for testing are shown in Table 1.
表1园林小菊品种信息表Table 1 Garden Chrysanthemum species information table
供试材料于2021年5月定植于青海省农林科学院园艺所试验田，种植株距为40cm，行距为40cm，随机排列，采取相同的常规水肥管理方法。试验选择生长健壮、无病虫害的母株并剪取8～10cm的顶梢为插穗，留上部2～3片叶，扦插于50孔穴盘中，共设2个温度处理：昼/夜15℃/15℃和25℃/20℃，将材料分别置于不同温度条件下，其中以25℃作为对照，每个处理50根插穗。The test materials were planted in the experimental field of the Horticulture Institute of Qinghai Academy of Agriculture and Forestry Sciences in May 2021. The planting spacing was 40cm, the row spacing was 40cm, arranged randomly, and the same conventional water and fertilizer management methods were adopted. In the experiment, a mother plant with strong growth and no pests and diseases was selected, and the top tip of 8-10 cm was cut off as cuttings, and the upper 2-3 leaves were left, and the cuttings were placed in a 50-hole hole tray. 15°C and 25°C/20°C, the materials were placed under different temperature conditions, 25°C was used as the control, and 50 cuttings were treated for each treatment.
耐低温园林小菊品种的筛选，包含以下步骤：The screening of low temperature-resistant garden chrysanthemum varieties comprises the following steps:
(1)园林小菊根总表面积和根尖数的测定：(1) Determination of the total surface area of the roots of Chrysanthemum chrysanthemum and the number of root tips:
在园林小菊生长期，通过根系表型分析系统GXY-A测定其根总表面积和其根尖数，根总表面积以cm2为单位，根尖数以根为单位。During the growth period of Chrysanthemum chrysanthemum, the total root surface area and the number of root tips were measured by the root phenotype analysis system GXY-A. The total root surface area is in cm 2 , and the root tip number is in root.
(2)园林小菊总叶绿素含量的测定：在园林小菊生长期，测定其叶片中的总叶绿素含量：选取各品种在不同温度处理下的顶梢，洗净顶梢表面污物，剪碎混匀后，各称取剪碎鲜样0.1g，共3份，分别置于10mL离心管中。每个离心管中加入5mL 95％的乙醇，使样品完全浸泡在乙醇溶液中。置于暗处浸泡24h后，将叶绿素提取液倒入比色皿中，以95％乙醇溶液作为对照并进行调零，测定波长665nm和649nm下的吸光度。按公式计算出叶绿素a、叶绿素b和总叶绿素浓度，以此获得组织中色素的含量。(叶绿素a浓度＝13.95A665-6.88A649；叶绿素b浓度＝24.96A649-7.32A665；总叶绿素浓度＝叶绿素a浓度+叶绿素b浓度；叶绿体色素的含量＝(色素浓度*提取液体积)/样品鲜重)(2) Determination of the total chlorophyll content of the garden chrysanthemum: in the growth period of the garden chrysanthemum, measure the total chlorophyll content in its leaves: select the top tips of each variety treated at different temperatures, wash the surface dirt on the top tips, cut them into pieces and mix them evenly Afterwards, weigh 0.1 g of the chopped fresh samples, 3 parts in total, and place them in 10 mL centrifuge tubes. Add 5 mL of 95% ethanol to each centrifuge tube to completely immerse the sample in the ethanol solution. After soaking in a dark place for 24 hours, pour the chlorophyll extract into a cuvette, use 95% ethanol solution as a control and perform zero adjustment, and measure the absorbance at wavelengths of 665nm and 649nm. Calculate the concentration of chlorophyll a, chlorophyll b and total chlorophyll according to the formula, so as to obtain the content of pigment in the tissue. (Chlorophyll a concentration=13.95A 665-6.88A 649 ; Chlorophyll b concentration= 24.96A 649-7.32A 665 ; Total chlorophyll concentration=Chlorophyll a concentration+Chlorophyll b concentration; The content of chlorophyll pigment=(pigment concentration*extract volume) /sample fresh weight)
(3)计算步骤(1)和步骤(2)测定的指标的耐低温系数α：根总表面积耐低温系数X1＝(胁迫后测定值/对照测定值)*100％；总叶绿素含量耐低温系数X2＝(胁迫后测定值/对照测定值)*100％；根尖数耐低温系数X3＝(胁迫后测定值/对照测定值)*100％。(3) Calculate the low temperature resistance coefficient α of the index measured in step (1) and step (2): root total surface area low temperature resistance coefficient X 1 =(measured value after stress/contrast measured value)*100%; total chlorophyll content is resistant to low temperature Coefficient X 2 =(measured value after stress/measured value of control)*100%; low temperature resistance coefficient X 3 of root tip number=(measured value after stress/measured value of control)*100%.
(4)园林小菊低温适应性评价：通过以园林小菊各单项指标的耐低温系数作为自变量建立的最优回归方程进行低温适应性评价，按照以下公式计算：(4) Evaluation of low temperature adaptability of garden chrysanthemum: the low temperature adaptability evaluation is carried out through the optimal regression equation established with the low temperature resistance coefficient of each individual index of garden chrysanthemum as an independent variable, and calculated according to the following formula:
耐低温性综合评价值＝0.220+0.090X1+0.580X2+0.095X3。Comprehensive evaluation value of low temperature resistance = 0.220+0.090X 1 +0.580X 2 +0.095X 3 .
园林小菊耐低温性状筛选结果如表2所示，耐低温性综合评价值评分(D值)越高，表明材料的耐低温性越强，其中‘金陵潋滟’的D值最大，表明该品种耐低温性最强，经验证其评分排序与实际耐低温性相符(孙东雷等，2021)，其中‘金陵潋滟’、‘金陵红荷’为一类，属高度耐低温类型，是在10℃～25℃的温度下扦插能够生长的品种。Table 2 shows the screening results of the low temperature resistance traits of Chrysanthemum chrysanthemum. The higher the comprehensive evaluation value (D value) of low temperature resistance, the stronger the low temperature resistance of the material. Among them, the D value of 'Jinling Lianyan' is the largest, indicating that the variety It has the strongest low temperature resistance, and it has been verified that its scoring order is consistent with the actual low temperature resistance (Sun Donglei et al., 2021). Among them, 'Jinling Brilliant' and 'Jinling Honghe' belong to the type of high low temperature resistance, and are the most resistant to low temperature at 10°C. A variety that can be grown by cuttings at a temperature of ~25°C.
表2园林小菊耐低温性综合评价值结果Table 2 The results of the comprehensive evaluation value of low temperature resistance of garden small chrysanthemum
本申请发明人还研究了不同温度下6个园林小菊的形态差异，进一步验证了本申请所述方案的可靠性。The inventor of the present application also studied the morphological differences of 6 small garden chrysanthemums at different temperatures, further verifying the reliability of the scheme described in the present application.
实施例1的六个田园小菊品种分别选择生长健壮、无病虫害的母株并剪取8～10cm的顶梢为插穗，留上部2～3片叶，扦插于50孔穴盘中，共设4个温度处理：5℃、10℃、15℃、25℃。将材料分别置于不同温度条件下，每个处理50根插穗，分别于扦插后28d进行采样，观察生根发育情况。For the six garden Chrysanthemum varieties of Example 1, select the mother plants that are robust in growth and free from diseases and insect pests, and cut the top shoots of 8 to 10 cm as cuttings, leave the upper 2 to 3 leaves, and cut them in 50 hole trays, with 4 in total. Temperature treatment: 5°C, 10°C, 15°C, 25°C. The materials were placed under different temperature conditions, and 50 cuttings were treated for each treatment, and samples were taken 28 days after cutting to observe the rooting development.
结果发现，各品种在正常培养下(25℃)根系均能正常生长；15℃处理下除‘金陵礼花’外各品种均生根；在10℃下仅有‘金陵红荷’、‘金陵潋滟’有根系产生，其余品种均未生根；这也与实施例1所述方法筛选的结果一致，进一步证明本申请所述方法的可靠性。It was found that the roots of all varieties could grow normally under normal culture (25°C); under the treatment of 15°C, all varieties except 'Jinling Lihua' took root; at 10°C, only 'Jinlinghonghe' and 'Jinling 'A root system is produced, and all the other varieties do not take root; this is also consistent with the result of the method screening described in Example 1, further proving the reliability of the method described in the present application.
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