Determination of Chlorophyll Concentration in In Vitro Peanut Leaves

IF 2.2

Current protocols Pub Date : 2025-05-22 DOI:10.1002/cpz1.70150

Paola C. Faustinelli, Alicia N. Massa, Néstor W. Soria, Eliana López Colomba, Paola A. Suárez, Marshall C. Lamb

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Abstract

The detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase (PDS) gene, which is related to chlorophyll biosynthesis. Various instruments and protocols are available for measuring chlorophyll; however, a minimum leaf area is required for precise quantifications. In vitro plant regeneration often requires several months to produce enough leaf tissue for chlorophyll content analysis. In this study, we optimized an existing chlorophyll quantification method by using a minimal amount of in vitro leaf tissue (<2.0 mg) and a microvolume of chlorophyll extraction solution (3 µl) to quantify chlorophyll a, chlorophyll b, and total chlorophyll concentration using a UV-Vis spectrophotometer. This method will serve as a basic protocol for evaluating gene editing efficiency by correlating degrees of albinism (pigment content) with changes in the PDS gene sequence. Published 2025. This article is a U.S. Government work and is in the public domain in the USA.

Basic Protocol: Determination of chlorophyll content in in vitro peanut leaves

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花生离体叶片叶绿素浓度的测定

叶绿素的检测是在产生与叶绿素生物合成有关的植物烯去饱和酶（phytoene desaturase， PDS）基因突变时评估基因编辑效率的一种便捷方法。各种仪器和方案可用于测量叶绿素；然而，精确定量需要最小叶面积。离体植株再生通常需要几个月的时间来产生足够的叶片组织以进行叶绿素含量分析。在本研究中，我们优化了现有的叶绿素定量方法，采用微量的离体叶片组织（2.0 mg）和微体积的叶绿素提取液（3µl），利用紫外可见分光光度计定量叶绿素a、叶绿素b和总叶绿素浓度。该方法将作为通过将白化程度（色素含量）与PDS基因序列变化相关联来评估基因编辑效率的基本方案。2025年出版。这篇文章是美国政府的作品，在美国属于公有领域。基本方案：离体花生叶片叶绿素含量的测定

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来源期刊

Current protocols

CiteScore

4.00

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0.00%

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