CN109580493A - 一种快速检测对节白蜡批量种子质量的方法 - Google Patents
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Abstract
本发明提供了一种快速检测对节白蜡批量种子质量的方法,主要包括以下步骤:将待测的批量对节白蜡种子平铺在黑色平面上,厚度为3‑6cm;使用地物光谱仪采集种子的反射光谱数据;对所述的反射光谱数据取平均值,剔除干扰较大的数据,获得原始谱图,从谱图的倾斜度即可快速判断批量的种子质量。该方法利用种子的反射光谱数据在700‑1050nm范围内具有良好的线性关系,建立批量对节白蜡种子的质量与近红外反射光谱之间的联系,为批量种子质量的判断提供依据,解决现有检测方法费时、费力、费用高等问题,在实际的生产生活中具有重大意义。
Description
技术领域
本发明涉及种子检测技术领域,具体涉及一种快速检测对节白蜡批量种子质量的方法。
背景技术
对节白蜡是木犀科白蜡属(梣属)一新种,全球范围内仅湖北特产。种子成熟后悬挂枝头,延至第2年春次第脱落,经1年休眠后才能出土萌发,1990年对节白蜡被正式确定为中国第2批珍惜濒危保护植物。该树种生长缓慢,枝叶浓密,树形优美,易于造型加工,是园林、盆景、根雕家族中的极品。此外,在水土保持、放养白蜡虫及植物地理学研究等方面,都具有很强的应用价值。
目前种子繁殖仍然是白蜡树属树种繁殖的主要方式。但对节白蜡具有深休眠特性,正常情况需要两冬一夏才能发芽,而且种子储存时外面还包裹有果皮,因此对于陈年种子很难通过外观进行确定。即使同一批次种子,在贮藏过程中,也会发生虫蛀、霉烂以及自身活力降低等问题,需要随时进行检验。对于休眠时间长的种子,即使通过刘雅琪等人的方法(去掉果皮,并去除胚根端被覆组织+激素处理),也需要三个月的时间才能判断种子发芽能力,对于快速判断批量种子活力远远不够。专利CN102960096B基于近红外光谱数据对水稻单籽粒活力进行无损检测,结合种子活力,建立一种活力与近红外光谱数据之间的联系,但存在检测难度大,数据庞大,处理困难等问题,对于批量种子总体质量,难以进行有效判断。而实际生产中,批量种子的质量判断具有更加重要的应用价值。
本发明通过对多批次种子的地物光谱仪数据分析,发现所得光谱数据在700-1050nm范围内具有良好的线性关系,且与种子的质量存在联系,因此比较起来非常方便,不需要繁杂的数据处理,大大提高了批量种子质量的判断效率。
发明内容
本发明要解决的技术问题是,以批量对节白蜡种子为研究对象,使用便携式地物光谱仪测定其反射光谱,建立批量对节白蜡种子的质量与近红外反射光谱之间的联系,为批量种子质量的判断提供依据,解决现有检测方法费时、费力、费用高等问题。
为实现上述目的,本发明提供了一种快速检测对节白蜡批量种子质量的方法,包括以下步骤:
(1)将待测的批量对节白蜡种子平铺在黑色平面上,厚度为3-6cm;
(2)使用地物光谱仪采集步骤(1)所述种子的反射光谱数据;
(3)对所述的反射光谱数据取平均值,剔除干扰较大的数据,获得原始谱图,从谱图的倾斜度即可快速判断批量的种子质量。
优选的,步骤(1)中,对节白蜡的种子带有果皮,每批量的种子数量为1000-20000个。
优选的,地物光谱仪的测定波段值为350-1050nm,采样间隔为0.5nm,光谱分辨率6nm。
优选的,步骤(2)所述地物光谱仪采集方法包括以下步骤:将地物光谱仪的探头平行置于种子上方,距离为5cm,选取平面不同位置多次采集。
探头与种子的距离为5cm时,信号采集面积约为25cm2,选择平面上不同的位置采集数据,根据种子平铺的面积设置采集次数,采集次数至少一次全面覆盖种子平铺面积。
优选的,步骤(c)所述剔除干扰较大的数据是剔除350-682nm的光谱数据。
本发明的有益效果:(1)地物光谱仪采集的是可见光波段(350-700nm)和短波近红外区(700-1050nm)的物质反射光谱,采集的数据少,但精确度更高,处理相对简单,大大提高了检测效率。(2)可用于批量对节白蜡种子质量的检测,通过与已知数据的比较,可以对批量种子活力迅速做出判断。(3)实现对不同批次种子质量的快速评价。(4)检测过程迅速,且不损害种子,同样适用于其他休眠期长的种子。
附图说明
图1为实施例1中不同批次对节白蜡种子的反射光谱图。
图2为实施例1中不同批次对节白蜡种子在682nm-1050nm的反射光谱图。
具体实施方式
下面将结合实施例对本发明做进一步的描述,但并不是限制本发明的范围,仅作示例说明。
实施例1:多批次对节白蜡种子的测定
选取3个批次的种子,每批次种子有1000个,将种子平铺在黑色桌面上,厚度为6cm。将地物光谱仪的探头平行置于种子上方,距离为5cm,选取平面不同位置多次采集,每批次样品采集6次,取数据的平均值,获得每批次种子的反射光谱图(图1)。从图1中可以看到,原始数据中含有大量噪声,这些噪声会影响地物光谱特征提取和识别精度,从而降低数据定量分析技术的有效性。噪声干扰除了一般图谱中的随机点状噪声外,高光谱图像中还受到严重的条带噪声干扰。通过对原始数据的分析,直接剔除噪声光谱,选取无噪声干扰的特定波长的反射光谱,从而便于对光谱数据进行分析,因此剔除350-682nm的光谱数据,重新作图(图2)。由图2可知,每条谱线在682nm至1050nm范围内,均呈现良好的线性关系。
根据谱线的倾斜度进行判断,倾斜度越高批量种子质量越低。因此批次3的种子质量最好,批次1次之,批次2是3个批次里质量最差的。
实施例2:可靠性检验
(1)腐烂率检测
取实施例1中检测后的对节白蜡种子,统计发霉腐烂的种子。检测方法为:人工去除对节白蜡种子的果皮,去掉虫蛀和霉烂的种子,然后通过手工的方式,对种子两端略施压力,如果种子断为两节,则为无效种子,活力基本丧失,如果比较坚硬,不易折断,则为高质量种子;虫蛀、霉烂以及无效种子计为发霉腐烂的种子。统计结果见表1.
表1:不同批次种子的腐烂率
批次 | 1 | 2 | 3 |
腐烂率 | 16% | 33% | 11% |
(2)生理生化指标测定
一般来讲,具有正常生理活性的种子能够在干燥条件下保持一定时间的活力,但是随着贮存时间的延长、贮存条件的不当、内外环境的胁迫,种子劣变老化将不可逆、不可避免地发生。种子老化或者劣变是一个逐步发生并且涉及众多复杂生理生化反应的过程。这体现在伴随着膜脂过氧化,保护酶钝化,蛋白质降解,相关基因表达紊乱以及核酸降解等生理生化反应,老化过程表现出种子发芽率、生活力、质膜完整性降低,内部保护酶系统活性减弱,有毒有害物质累积等特点。
研究表明老化种子的发芽势、发芽率、发芽指数、活力指数,相关保护酶活力均随老化时间的延长,整体呈下降趋势;种子浸出液电导率、膜脂过氧化产物——丙二醛(MDA)含量和可溶性糖含量均随老化时间的延长而升高,与种子的各项活力指标呈负相关。而且MDA是通过结合过氧化氢酶(CAT)、超氧化物歧化酶(SOD)等保护酶,致使保护酶空间构象改变,导致自由基累积而促成膜脂发生过氧化。
因此,选取通过腐烂检测的外观质量较好的种子,进行相关生理生化指标的测定:可溶性糖采用蒽酮比色法;可溶性蛋白采用BCA法(南京建成,A045-3);SOD利用超氧化物歧化酶测定试剂盒(南京建成,AOO1-4)测定;过氧化物酶(POD)采用过氧化物酶试剂盒(南京建成,A084-3)测定;CAT测定采用紫外吸收法。检测结果见表2。
表2:不同批次种子的生理生化指标
指标 | 批次1 | 批次2 | 批次3 |
POD | 1539.05 | 1838.85 | 1900.10 |
SOD | 112.52 | 114.38 | 114.29 |
CAT | 14.70 | 13.80 | 10.25 |
电导率 | 11.55 | 11.32 | 4.65 |
可溶性蛋白(μg/g) | 3.68 | 7.49 | 12.26 |
可溶性糖 | 1.86a | 2.00 | 1.55 |
结果表明,对节白蜡种子比较耐贮藏,挑选出的种子在POD、SOD、CAT等方面均无显著性差异,电导率和与种子活性关系较密切,电导率越低,表明种子活性越高。可溶性蛋白与种子活力成反比,表明随着贮藏时间的延长,可溶性蛋白含量降低,种子活力下降。可溶性糖含量随着贮藏时间的延长,含量会逐渐增大,含量越高,表明种子活性越差。
(3)发芽率检测:
对每批次种子,随机选择100粒,重复三次,进行发芽率检测。
采用长江大学刘雅琪等人的方法检测发芽率,首先去除对节白蜡种子的果皮和种皮,然后对对节白蜡种子进行切除胚根+赤霉素处理,置于4℃,低温层积,60天后,统计发芽率。检测结果见表3:
表3:不同批次种子的发芽率
通过对3个批次种子的腐烂率、发芽率以及多种生理生化指标的检测,与实施例1的检测结果是一致的,因此本发明提供的方法是可靠的。
对于本领域的技术人员来说,可根据以上描述的技术方案以及构思,做出其它各种相应的改变以及变形,而所有的这些改变以及变形都应该属于本发明权利要求的保护范围之内。
Claims (5)
1.一种快速检测对节白蜡批量种子质量的方法,其特征在于:包括以下步骤:
a)将待测的批量对节白蜡种子平铺在黑色平面上,厚度为3-6cm;
b)使用地物光谱仪采集步骤a)所述种子的反射光谱数据;
c)对所述的反射光谱数据取平均值,剔除干扰较大的数据,获得谱图,从谱图的倾斜度即可快速判断批量的种子质量。
2.根据权利要求1所述一种快速检测对节白蜡批量种子质量的方法,其特征在于:步骤a)所述对节白蜡的种子带有果皮,每批量种子有1000-20000个。
3.根据权利要求1所述一种快速检测对节白蜡批量种子质量的方法,其特征在于:步骤b)所述地物光谱仪的测定波段值为350-1050nm,采样间隔为0.5nm,光谱分辨率6nm。
4.根据权利要求1所述一种快速检测对节白蜡批量种子质量的方法,其特征在于:步骤b)所述地物光谱仪采集方法包括以下步骤:将地物光谱仪的探头平行置于种子上方,距离为5cm,选取不同位置多次采集。
5.根据权利要求1所述一种快速检测对节白蜡批量种子质量的方法,其特征在于:步骤c)所述剔除干扰较大的数据是剔除350-682nm的光谱数据。
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