PCRmax Botrytis cinerea Instruction Manual

Species-specific fragment

Botrytis cinerea

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Botryotinia cinerea is a spore forming fungus of the Botryotinia genus with a haploid DNA
genome of approximately 38 Mb. The asexual form is called Botrytis cinerea, while the
sexual form is Botryotinia cinerea (also known as Botryotinia fuckeliana), although they are
the same organism. The anamorphic form blights many fruit crops causing rot.

The organism over-winters in one of two ways depending on the conditions. The first is as
mycelia – the mass of branching hyphae living on decaying plant matter; while the second
is as sclerotia – which is where the mycelia harden and compact, then detaches from the
fungi. When conditions improve in the spring, new growth begins with the production of
condiophores bearing conidia. These conidia detach and are dispersed by wind to attack
new hosts. Initial infection usually occurs in a previously damaged tissue that has
increased susceptibility although seedlings are also vulnerable. New mycelia then invade
the tissue causing cells death which results in tissue softening and eventually decay.
These mycelia ultimately produce new condiophores and the cycle continues.
Fungicide can often be used to prevent B. cinerea infection. Preventing the spread of this
disease usually involves removing infected plants or parts of the plant during dry
conditions.

In grapes – the most commercially affected plant – the fungus causes two conditions: grey
rot, in wet or humid conditions which results in the loss of infected bunches; and noble rot,
in dry conditions which can result in sweetening the grapes. The fungus also infects many
other plants including vegetables and fruit. In these plants it can cause leaf spots, shoot or
bud blights, bud blats or cankers depending on the specific part of the plant that is infected.

Introduction to Botrytis cinerea

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MIN

MAX

The intergenic spacer region has previously been identified as a highly specific market for
Botryotinia cinerea (Plant Physiol Biochem. 2005 Sep;43(9):890-9) and the primers have
100% homology with all reference sequences for Botryotinia cinerea in the NCBI
database (DQ000001.1, AY674786.1, AY694147.1, AY694146.1, AJ422103.1,
AJ539088.1).

The PCR Max qPCR Kit for Botrytis cinerea (B.cinerea) genomes is designed for the in
vitro quantification of B.cinerea genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the B.cinerea genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
B.cinerea sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• B.cinerea specific primer/probe mix (150 reactions BROWN)

FAM labelled

• B.cinerea positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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